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1
Ou-Yang, Huan, Shinn-Chih Wu, Li-Ying Sung, Shiao-Hsuan Yang, Shang-Hsun Yang, Kowit-Yu Chong y Chuan-Mu Chen. "STAT3 Is an Upstream Regulator of Granzyme G in the Maternal-To-Zygotic Transition of Mouse Embryos". International Journal of Molecular Sciences 22, n.º 1 (5 de enero de 2021): 460. http://dx.doi.org/10.3390/ijms22010460.
Resumen
The maternal-to-zygotic transition (MZT), which controls maternal signaling to synthesize zygotic gene products, promotes the preimplantation development of mouse zygotes to the two-cell stage. Our previous study reported that mouse granzyme g (Gzmg), a serine-type protease, is required for the MZT. In this study, we further identified the maternal factors that regulate the Gzmg promoter activity in the zygote to the two-cell stage of mouse embryos. A full-length Gzmg promoter from mouse genomic DNA, FL-pGzmg (−1696~+28 nt), was cloned, and four deletion constructs of this Gzmg promoter, Δ1-pGzmg (−1369~+28 nt), Δ2-pGzmg (−939~+28 nt), Δ3-pGzmg (−711~+28 nt) and Δ4-pGzmg (−417~+28 nt), were subsequently generated. Different-sized Gzmg promoters were used to perform promoter assays of mouse zygotes and two-cell stage embryos. The results showed that Δ4-pGzmg promoted the highest expression level of the enhanced green fluorescent protein (EGFP) reporter in the zygotes and two-cell embryos. The data suggested that time-specific transcription factors upregulated Gzmg by binding cis-elements in the −417~+28-nt Gzmg promoter region. According to the results of the promoter assay, the transcription factor binding sites were predicted and analyzed with the JASPAR database, and two transcription factors, signal transducer and activator of transcription 3 (STAT3) and GA-binding protein alpha (GABPα), were identified. Furthermore, STAT3 and GABPα are expressed and located in zygote pronuclei and two-cell nuclei were confirmed by immunofluorescence staining; however, only STAT3 was recruited to the mouse zygote pronuclei and two-cell nuclei injected with the Δ4-pGzmg reporter construct. These data indicated that STAT3 is a maternal transcription factor and may upregulate Gzmg to promote the MZT. Furthermore, treatment with a STAT3 inhibitor, S3I-201, caused mouse embryonic arrest at the zygote and two-cell stages. These results suggest that STAT3, a maternal protein, is a critical transcription factor and regulates Gzmg transcription activity in preimplantation mouse embryos. It plays an important role in the maternal-to-zygotic transition during early embryonic development.
2
Chiang, Jeffrey, Wai Lim Ku, Kairong Cui, Keji Zhao y Richard J. Hodes. "The use of alternative promoters in T cell development". Journal of Immunology 200, n.º 1_Supplement (1 de mayo de 2018): 165.20. http://dx.doi.org/10.4049/jimmunol.200.supp.165.20.
Resumen
Abstract Many mammalian genes, including a number involved in immune development and function, use multiple promoters encoding expression of the same protein product. To understand the function of alternative promotors during T cell development, we carried out genome-wide RNA-Seq analysis of DN, DP, CD4 and CD8 thymocytes. 26% (2469/9595) of genes expressed by thymocytes used two or more alternative promotors at some point in T cell development. However, only 3.2% (80/2469) of the genes which utilized two or more promoters showed the difference of promotor activity among different T cell developmental stages which were validated by selected representative genes with qPCR assay. To analyze mechanisms of alternative promoter regulation, DNase hypersensitivity sites were mapped, and ChIP-Seq with anti-H3K4me3 antibodies was performed in the same thymic subpopulations. To determine the in vivo functional role of alternative promotors for T cell development, several genes were selected for study by deletion of individual promoters by CRISPR technology. We have initially generated mice with deletion of distal or proximal promotor of the lck gene and demonstrated distinct functional roles of the two promoters: the proximal lck promoter is critical for early stages of thymic T cell development while the distal promoter was necessary for normal T cell activation. Ongoing studies will characterize more broadly the functional role of alternative promoter expression in thymocyte development, and the mechanisms mediating selective promoter activity.
3
Zhu, Guangwei, Qiang Huang, Wei Zheng, Yongjian Huang, Jin Hua, Shugang Yang, Jinfu Zhuang et al. "LPS Upregulated VEGFR-3 Expression Promote Migration and Invasion in Colorectal Cancer via a Mechanism of Increased NF-κB Binding to the Promoter of VEGFR-3". Cellular Physiology and Biochemistry 39, n.º 5 (2016): 1665–78. http://dx.doi.org/10.1159/000447868.
Resumen
Background and Aim: Lipopolysaccharide(LPS) could promote the progression of colorectal cancer, but the specific regulatory mechanisms are largely unknown. So, this study aim to clarify the mechanisms that LPS upregulated VEGFR-3, which promotes colorectal cancer cells migration and invasion with a mechanism of increased NF-κB bind to the promoter of VEGFR-3. Methods: The present study examined the VEGFR-3 expression in colorectal cancer tissues and analyzed the relationship between the VEGFR-3 expression with clinical parameters. PCR, Western blot, CCK-8, colone formation assay, and Transwell assay detected that LPS promoted the migration and invasion and the role of VEGFR-3 in the process of colorectal carcinoma in vitro. Used the methods of promoter analysis, EMSA assay and ChIP assay to explore the mechanisms LPS increased the expression of VEGFR-3. Results: VEGFR-3 was significantly high expression in the colorectal cancer tissues. And the high expression was associated with the TNM stage and lymph node metastasis of colorectal cancer. LPS could promote the migration and invasion, which could be blocked by the neutralizing antibody IgG of VEGFR-3. And found that -159 nt to +65 nt was the crucial region of VEGFR-3 promoter. And detected that the NF-κB was important transcription factor for the VEGFR-3 promoter. And LPS could increase NF-κB binding to VEGFR-3 promoter and upregulated the expression of VEGFR-3 to exert biological functions. Conclusion: We have elucidated the relationship between LPS and the VEGFR-3 expression and revealed that VEGFR-3 play very important role in the process of LPS promoting the migration and invasion of colorectal cancer cells. Further illuminated the mechanism that LPS upregulated VEGFR-3 expression via increased NF-κB bind to the promoter of VEGFR-3.
4
He, Yang, Tao Zhao, Fang Chen, Changchun Song, Chongchao Zhong y Zhi Luo. "Functional Analysis of the Promoter Regions of Two Apoptosis-Related Genes (Bcl-2 and Cycs) and Their Regulation by Zn in Yellow Catfish". International Journal of Molecular Sciences 22, n.º 12 (11 de junio de 2021): 6291. http://dx.doi.org/10.3390/ijms22126291.
Resumen
B-cell lymphoma 2 (Bcl-2) and cytochrome c (Cycs) are two important proteins relevant to cellular apoptosis. In this study, we characterized the functions of the promoter regions of two apoptosis-related genes, Bcl-2 and Cycs, in yellow catfish Pelteobagrus fulvidraco. We obtained a 1989 bp Bcl-2 promoter and an 1830 bp Cycs promoter and predicted several key transcription factor binding sites (TFBSs) on the promoters, such as Kruppel-like factor 4 (KLF4), signal transducer and activator of transcription factor 3 (STAT3), forkhead box O (FOXO), metal-responsive element (MRE) and hepatocyte nuclear factor 1α (HNF-1α). Zinc (Zn) increased the activities of the Bcl-2 promoter but decreased the activities of the Cycs promoter. Metal-responsive transcription factor 1 (MTF-1) and HNF-1α directly bound with Bcl-2 and Cycs promoters, and they positively regulated the activity of the Bcl-2 promoter but negatively regulated the activity of the Cycs promoter. Zn promoted the binding ability of HNF-1α to the Bcl-2 promoter but decreased its binding ability to the Cycs promoter. However, Zn had no significant effect on the binding capability of MTF-1 to the regions of Bcl-2 and Cycs promoters. Zn upregulated the mRNA and total protein expression of Bcl-2 but downregulated the mRNA and total protein expression of Cycs. At the same time, Annexin V–FITC/PI staining showed that Zn significantly reduced the apoptosis of primary hepatocytes. For the first time, our study provides evidence for the MRE and HNF-1α response elements on the Bcl-2 and Cycs promoters, offering new insight into the mechanism by which Zn affects apoptosis in vertebrates.
5
Tang, Hongting, Yanling Wu, Jiliang Deng, Nanzhu Chen, Zhaohui Zheng, Yongjun Wei, Xiaozhou Luo y Jay D. Keasling. "Promoter Architecture and Promoter Engineering in Saccharomyces cerevisiae". Metabolites 10, n.º 8 (6 de agosto de 2020): 320. http://dx.doi.org/10.3390/metabo10080320.
Resumen
Promoters play an essential role in the regulation of gene expression for fine-tuning genetic circuits and metabolic pathways in Saccharomyces cerevisiae (S. cerevisiae). However, native promoters in S. cerevisiae have several limitations which hinder their applications in metabolic engineering. These limitations include an inadequate number of well-characterized promoters, poor dynamic range, and insufficient orthogonality to endogenous regulations. Therefore, it is necessary to perform promoter engineering to create synthetic promoters with better properties. Here, we review recent advances related to promoter architecture, promoter engineering and synthetic promoter applications in S. cerevisiae. We also provide a perspective of future directions in this field with an emphasis on the recent advances of machine learning based promoter designs.
6
O’Callaghan, Chris, Da Lin y Thomas K. Hiron. "Intragenic transcriptional interference regulates the human immune ligand MICA." Journal of Immunology 200, n.º 1_Supplement (1 de mayo de 2018): 109.23. http://dx.doi.org/10.4049/jimmunol.200.supp.109.23.
Resumen
Abstract Regulation of MICA expression is incompletely understood, but human MICA can be upregulated in cancer cells, virus-infected cells and rapidly proliferating cells. Binding of MICA to the activating NKG2D receptor on cytotoxic immune cells promotes elimination of the cell expressing MICA. We noted that MICA has tandem promoters that drive overlapping forward transcription. We show that the MICA gene contains a conserved upstream promoter that expresses a non coding transcript. Transcription from the upstream promoter represses transcription from the standard downstream MICA promoter in cis through transcriptional interference. The effect of transcriptional interference depends on the strength of transcription from the upstream promoter and quantitative studies show that it is described by a simple reciprocal repressor function. The time course of transcriptional interference coincides with recruitment at the standard downstream promoter of factors involved in nucleosomal remodeling during transcription. Transcriptional interference is demonstrated in the regulation of MICA expression by the physiological inputs interferon-γ and interleukin-4, that both act through regulatory DNA elements in the upstream promoter. These findings have significant implications for the understanding of MICA expression. Transcription factors activating the downstream promoter will upregulate MICA expression, whereas transcription factors activating the upstream promoter will downregulated MICA expression. A genome-wide analysis indicates that transcriptional interference between tandem intragenic promoters may be involved in regulating the expression of multiple other human genes.
7
Tasfy, Sara Faiz Hanna, Noor Asmawati Mohd Zabidi y Duvvuri Subbarao. "Effects of Cu and K Promoters on the Catalytic Performance of Iron-Based Nanocatalyst for Fischer-Tropsch Synthesis". Advanced Materials Research 832 (noviembre de 2013): 15–20. http://dx.doi.org/10.4028/www.scientific.net/amr.832.15.
Resumen
Iron-based nanocatalyst was prepared via impregnation method on SiO2 support. The effects of promoters, namely, K and Cu, on the physical properties and catalytic performance in FTS have been investigated. The FTS performance of the synthesized nanocatalysts was examined in a fixed-bed microreactor at temperature of 523K, atmospheric pressure, 1.5 reactant ratio (H2/CO) and space velocity of 3L/g-cat.h. In FTS reaction, Cu promoter resulted in a lower CO conversion and C5+ hydrocarbons selectivity but higher selectivity to the lighter hydrocarbons (C1-C4) comparedto those obtained using the K promoter. Higher CO conversion (28.9%) and C5+ hydrocarbons selectivity (54.4%) were obtained using K as a promoter compared to that of Cu promoter. However, the K-promoted nanocatalyst resulted in a lower CO conversion but higher selectivity of the heavy hydrocarbons (C5+) compared to those obtained using the un-promoted nanocatalyst.
8
Yang, Hong, Chongchao Zhong, Xiaoying Tan, Guanghui Chen, Yang He, Shengzan Liu y Zhi Luo. "Transcriptional Responses of Copper-Transport-Related Genes ctr1, ctr2 and atox1 and Their Roles in the Regulation of Cu Homeostasis in Yellow Catfish Pelteobagrus fulvidraco". International Journal of Molecular Sciences 23, n.º 20 (13 de octubre de 2022): 12243. http://dx.doi.org/10.3390/ijms232012243.
Resumen
Here, we characterized the function of ctr1, ctr2 and atox1 promoters in yellow catfish Pelteobagrus fulvidraco, a common freshwater teleost in Asian countries. We obtained 1359 bp, 1842 bp and 1825 bp sequences of ctr1, ctr2 and atox1 promoters, and predicted key transcription factor binding sites on their promoters, including MRE, SREBP1, NRF2, KLF4 and STAT3. Cu differentially influenced the activities of ctr1, ctr2 and atox1 promoters from different regions. We found that the −326/−334 bp and −1232/−1240 bp locus in the atox1 promoter were functional NRF2 binding sites, which negatively controlled the activity of the atox1 promoter. The −91/−100 bp locus in the ctr1 promoter and −232/−241 bp and −699/−708 bp locus in the atox1 promoter were functional SREBP1 binding sites, which positively controlled the activities of ctr1 and atox1 promoters. Cu inhibited the NRF2 binding ability to the atox1 promoter, but promoted the SREBP1 binding ability to the ctr1 and atox1 promoters. Dietary Cu excess significantly down-regulated hepatic mRNA and total protein expression of CTR1, CTR2 and ATOX1 of yellow catfish, compared to the adequate dietary Cu group. The subcellular localization showed that CTR1 was mainly localized on the cell membrane, CTR2 in the cell membrane and the lysosome, and ATOX1 in the cytoplasm. In conclusion, we demonstrated the regulatory mechanism of three Cu transporters at the transcription levels, and found the functional NRF2 and SREBP1 response elements in ctr1, ctr2 and atox1 promoters, which provided new insights into their roles in the regulation of Cu homeostasis in fish.
9
Zhang, Chong, Huxia Gu, Dingrong Liu, Jing Fang y Yan Yang. "The Role of MRPL23 Antisense RNA 1 (MRPL23-AS1) in the Pre-Metastatic Microenvironment of Malignancy During the Process of Epithelial-Mesenchymal Transition". Journal of Biomaterials and Tissue Engineering 11, n.º 5 (1 de mayo de 2021): 864–71. http://dx.doi.org/10.1166/jbt.2021.2638.
Resumen
We aimed to explore MRPL23-AS1’s role in the pre-metastatic microenvironment of malignancy during epithelial-mesenchymal transition (EMT). Identification and verification of lncRNA-interacting proteins in salivary adenoid cystic carcinoma (SACC) cells were conducted via RNA-pulldown, silver staining, and Western blotting. RIP and RIP-seq were sequentially administered to verify the binding partners of lncRNA. CHIRP was performed to detect the promoter DNA in the downstream of lncRNA-protein complex. Ultimately CHIP-qPCR detected the effects of lncRNA on the binding degree of its interacting protein to the promoter DNA in the downstream genes and the methyla-tion level of histones in the promoter region. The exosomes secreted by different SACC cells were extracted from culture supernatant to measure lncRNA expression via qPCR. MRPL23-AS1 interacted with EZH2 protein and promoted EZH2 binding to E-cadherin gene promoter region along with the H3K27 methylation. MRPL23-AS1 could promote EMT of SACC cells and increase pulmonary vascular endothelial cells permeability via exosomes secretion. MRPL23-AS1 up-regulated VEGFA, while down-regulated E-cadherin and VE-cadherin in endothelial cells. Exosomes rich in MRPL23-AS1 could boost lung metastasis in vivo. MRPL23-AS1 inhibits E-cadherin level and promotes EMT of SACC cells, suggesting that it might be a biomarker and therapeutic target for lung cancer.
10
Cheng, Wenlong, Yongqiang Qi, Li Tian, Bing Wang, Wenhua Huang y Yongjun Chen. "Dicer promotes tumorigenesis by translocating to nucleus to promote SFRP1 promoter methylation in cholangiocarcinoma cells". Cell Death & Disease 8, n.º 2 (febrero de 2017): e2628-e2628. http://dx.doi.org/10.1038/cddis.2017.57.
11
Chen, Shu-Wei, Kun Wu, Wu-Hong Lv, Fang Chen, Chang-Chun Song y Zhi Luo. "Functional Analysis of Two Zinc (Zn) Transporters (ZIP3 and ZIP8) Promoters and Their Distinct Response to MTF1 and RREB1 in the Regulation of Zn Metabolism". International Journal of Molecular Sciences 21, n.º 17 (26 de agosto de 2020): 6135. http://dx.doi.org/10.3390/ijms21176135.
Resumen
ZIP (zinc-regulated transporters, iron-regulated transporter-like protein) family plays an important role in organism Zn balance. This research identified the promoter regions of ZIP3 and ZIP8, two members of ZIP family, from a freshwater teleost yellow catfish Pelteobagrus fulvidraco, characterized the binding sequences of the metal-responsive transcription factor-1 (MTF-1) and Ras responsive element binding protein 1 (RREB1) on their promoter regions. The present study cloned and obtained the 2027 bp of ZIP3 promoter and 1664 bp of ZIP8 promoter, and predicted several key elements on their promoters, such as the binding sites of CREB (cAMP-response element binding protein), KLF4 (Kruppel like factor 4), MTF-1 and RREB1. The sequence deletion from −361 bp to −895 bp down-regulated the luciferase activity of ZIP3 promoter, and the deletion from −897 bp to −1664 bp down-regulated the luciferase activity of ZIP8 promoter. Within different deletion plasmids, the relative luciferase activities of ZIP3 and ZIP8 promoters changes to Zn incubation in a Zn concentration-dependent manner. The site mutagenesis and EMSA (electrophoretic mobility shift assay) found that the −1327 bp/−1343 bp MTF-1 binding site and the −248 bp/−267 bp RREB1 binding site on the ZIP3 promoter, and the −1543 bp/−1557 bp MTF-1 binding site on the ZIP8 promoter are functional sites. Low Zn increased the binding capability between MTF-1 and its responsive site on the ZIP3 promoter, and high Zn increased the transcriptional activation ZIP3 by RREB1; Zn also promoted the binding ability between MTF-1 and its responsive element on the ZIP8 promoter. This study provides the first direct evidence for the response elements of MTF-1 and RREB1 on ZIP3 and MTF-1 on ZIP8 to Zn, which are very important for the evaluation of Zn nutrition and toxicity in vertebrates.
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DesJardins, E. y N. Hay. "Repeated CT elements bound by zinc finger proteins control the absolute and relative activities of the two principal human c-myc promoters". Molecular and Cellular Biology 13, n.º 9 (septiembre de 1993): 5710–24. http://dx.doi.org/10.1128/mcb.13.9.5710-5724.1993.
Resumen
Transcription of the human proto-oncogene c-myc is governed by two tandem principal promoters, termed P1 and P2. In general, the downstream promoter, P2, is predominant, which is in contrast to the promoter occlusion phenomenon usually observed in genes containing tandem promoters. A shift in human c-myc promoter usage has been observed in some tumor cells and in certain physiological conditions. However, the mechanisms that regulate promoter usage are not well understood. The present studies identify regulators which are required to promote transcription from both human c-myc promoters, P1 and P2, and have a role in determining their relative activities in vivo. A novel regulatory region located 101 bp upstream of P1 was characterized and contains five tandem repeats of the consensus sequence CCCTCCCC (CT element). The integrity of the region containing all five elements is required to promote transcription from P1 and for maximal activity from P2 in vivo. A single copy of this same element, designated CT-I2, also appears in an inverted orientation 53 bp upstream of the P2 transcription start site. This element has an inhibitory effect on P1 transcription and is required for P2 transcription. The transcription factor Sp1 was identified as the factor that binds specifically to the tandem CT elements upstream of P1 and to the CT-I2 element upstream of P2. In addition, the recently cloned zinc finger protein ZF87, or MAZ, was also able to bind these same elements in vitro. The five tandem CT elements can be functionally replaced by a heterologous enhancer that only in the absence of CT-I2 reverses the promoter usage, similar to what is observed in the translocated c-myc allele of Burkitt's lymphoma cells.
13
DesJardins, E. y N. Hay. "Repeated CT elements bound by zinc finger proteins control the absolute and relative activities of the two principal human c-myc promoters." Molecular and Cellular Biology 13, n.º 9 (septiembre de 1993): 5710–24. http://dx.doi.org/10.1128/mcb.13.9.5710.
Resumen
Transcription of the human proto-oncogene c-myc is governed by two tandem principal promoters, termed P1 and P2. In general, the downstream promoter, P2, is predominant, which is in contrast to the promoter occlusion phenomenon usually observed in genes containing tandem promoters. A shift in human c-myc promoter usage has been observed in some tumor cells and in certain physiological conditions. However, the mechanisms that regulate promoter usage are not well understood. The present studies identify regulators which are required to promote transcription from both human c-myc promoters, P1 and P2, and have a role in determining their relative activities in vivo. A novel regulatory region located 101 bp upstream of P1 was characterized and contains five tandem repeats of the consensus sequence CCCTCCCC (CT element). The integrity of the region containing all five elements is required to promote transcription from P1 and for maximal activity from P2 in vivo. A single copy of this same element, designated CT-I2, also appears in an inverted orientation 53 bp upstream of the P2 transcription start site. This element has an inhibitory effect on P1 transcription and is required for P2 transcription. The transcription factor Sp1 was identified as the factor that binds specifically to the tandem CT elements upstream of P1 and to the CT-I2 element upstream of P2. In addition, the recently cloned zinc finger protein ZF87, or MAZ, was also able to bind these same elements in vitro. The five tandem CT elements can be functionally replaced by a heterologous enhancer that only in the absence of CT-I2 reverses the promoter usage, similar to what is observed in the translocated c-myc allele of Burkitt's lymphoma cells.
14
Chiang, Jeffrey y Richard J. Hodes. "Long-range transcriptional effects and specific epigenetic marks in regulation of transcription from Lck proximal and distal promoters". Journal of Immunology 196, n.º 1_Supplement (1 de mayo de 2016): 128.17. http://dx.doi.org/10.4049/jimmunol.196.supp.128.17.
Resumen
Abstract Lck is a critical molecule for T cell development and activation. The T cell-specific expression of Lck is controlled by two distinct promoters separated by 15kb. The proximal promoter contributes the majority of Lck transcripts in immature thymic T cells, while the distal promoter expresses the majority of Lck transcripts in mature T cells. To study the regulation of these two promoters, promoter-mutated mouse models were generated by CRISPR technology, resulting in multiple mouse lines with distinct promoter deletions. The activity of the proximal promoter was reduced by 95% in the DP thymocytes of PROX[mut] mice with ~100bp deletion in the proximal promoter. Expression of distal-promoter-driven Lck mRNA was completely abrogated in immature and mature T cells in Lck distal promoter DIST[mut] mice with partial deletion of the distal promoter and exon 1. Distal-promoter-driven Lck mRNAs were not altered in PROX[mut] mice. However, proximal-promoter-driven Lck mRNAs were dramatically increased in the mature T cells of DIST[mut] mouse, suggesting that there is a long-range transcriptional interaction between the two promoters. To further understand the epigenetic influences in Lck gene expression, we analyzed histone modifications in the regions of Lck promoters. Our preliminary data showed the distal promoter was highly H3K9 acetylated in peripheral T cells but not immature T cells, and that this acetylation was decreased in DIST[mut] mice. Therefore, distal promoter H3K9 acetylation was correlated with promoter activity and may be involved in the developmental stage specificity of Lck promoters. The findings of this study on Lck promoters may provide important information on gene expression and its tissue-specificity.
15
Govindarajulu, Manjula, James M. Elmore, Thomas Fester y Christopher G. Taylor. "Evaluation of Constitutive Viral Promoters in Transgenic Soybean Roots and Nodules". Molecular Plant-Microbe Interactions® 21, n.º 8 (agosto de 2008): 1027–35. http://dx.doi.org/10.1094/mpmi-21-8-1027.
Resumen
The efficiency of β-glucuronidase (GUS) expression was evaluated with five viral promoters to identify the most suitable promoter or promoters for use in soybean hairy roots, including applications to study the symbiotic interaction with Bradyrhizobium japonicum. Levels of GUS activity were fluorimetrically and histochemically assayed when the GUS (uidA) gene was driven by the Cauliflower mosaic virus (CaMV) 35S promoter and enhanced 35S (E35S) promoter, the Cassava vein mosaic virus (CsVMV) promoter, the Figwort mosaic virus (FMV) promoter, and the Strawberry vein banding virus (SVBV2) promoter. We demonstrate that GUS activity was highest when driven by the FMV promoter and that the promoter activity of 35S and SVBV2 was significantly lower than that of the CsVMV and E35S promoters when tested in soybean hairy roots. In mature soybean root nodules, strong GUS activity was evident when the FMV, 35S, and CsVMV promoters were used. These results indicate that the FMV promoter facilitates the strong expression of target genes in soybean hairy roots and root nodules.
16
Cichocki, Frank, Rebecca J. Hanson, Todd Lenvik, Michelle Pitt, Valarie McCullar, Hongchuan Li, Stephen K. Anderson y Jeffrey S. Miller. "The transcription factor c-Myc enhances KIR gene transcription through direct binding to an upstream distal promoter element". Blood 113, n.º 14 (2 de abril de 2009): 3245–53. http://dx.doi.org/10.1182/blood-2008-07-166389.
Resumen
Abstract The killer cell immunoglobulin-like receptor (KIR) repertoire of natural killer (NK) cells determines their ability to detect infected or transformed target cells. Although epigenetic mechanisms play a role in KIR gene expression, work in the mouse suggests that other regulatory elements may be involved at specific stages of NK-cell development. Here we report the effects of the transcription factor c-Myc on KIR expression. c-Myc directly binds to, and promotes transcription from, a distal element identified upstream of most KIR genes. Binding of endogenous c-Myc to the distal promoter element is significantly enhanced upon interleukin-15 (IL-15) stimulation in peripheral blood NK cells and correlates with an increase in KIR transcription. In addition, the overexpression of c-Myc during NK-cell development promotes transcription from the distal promoter element and contributes to the overall transcription of multiple KIR genes. Our data demonstrate the significance of the 5′ promoter element upstream of the conventional KIR promoter region and support a model whereby IL-15 stimulates c-Myc binding at the distal KIR promoter during NK-cell development to promote KIR transcription. This finding provides a direct link between NK-cell activation signals and KIR expression required for acquisition of effector function during NK-cell education.
17
Arsenijevic, Slavica y Ljubisa Topisirovic. "Molecular analysis of mutatedLactobacillus acidophiluspromoter-like sequence P15". Canadian Journal of Microbiology 46, n.º 10 (1 de octubre de 2000): 938–45. http://dx.doi.org/10.1139/w00-077.
Resumen
The promoter-like sequence P15 that was previously cloned from the chromosome of Lactobacillus acidophilus ATCC 4356 is active in Lactobacillus reuteri, Lactobacillus plantarum, Lactobacillus acidophilus, and Escherichia coli, but not in Lactococcus lactis. N-methyl-N-nitroso-N-guanidine (MNNG) mutagenesis of P15 was used to select for a promoter active in L. lactis MG1363. Molecular analysis of the mutated promoter (designated P16) revealed a 90 bp deletion and a T[Formula: see text]A transversion. This deletion, in combination with the addition to the transversion, created a promoter with putative -35 and -10 hexamers identical to the consensus promoter sequence found in E. coli and Bacillus subtilis vegetative promoters. The activity of P16 was measured by its ability to promote chloramphenicol resistance in different bacteria when inserted in the promoter-probe plasmid pBV5030 (designated pLA16). The MIC of chloramphenicol in L. lactis, L. reuteri, L. plantarum, E. coli, and L. acidophilus harbouring pLA16 were 30, 170, 180, >500, and 3 µg/mL, respectively. This represents an increase in promoter activity compared to P15 in L. reuteri of 3-fold, in L. plantarum of 9-fold, and in E. coli of at least 2.5-fold, but a decrease in L. acidophilus of 7-fold.Key words: Lactobacillus acidophilus, promoter-like sequence, mutagenesis.
18
Bacha, Sardar, Nadia Iqbal, Aftab Bashir, Farah Deeba, Muhammad Asif y Joshua Yuan. "Evaluation of Cis-Regulatory Elements of Dicot ?-TbSt Promoter". JOURNAL OF MICROBIOLOGY AND MOLECULAR GENETICS 2, n.º 3 (31 de diciembre de 2021): 23–38. http://dx.doi.org/10.52700/jmmg.v2i3.35.
Resumen
The eukaryotic gene expression is controlled by a regulatory region called promoter. Many plant promoters have been characterized for regulatory motifs. There are three types of plant promoters i.e. inducible, constitutive and tissue specific on basis of regulatory motifs. Plant sources have been searched for isolation of strong promoters that are being utilized in molecular biology research. The researchers need to address IPR issues for utilizing the strong patented promoters for the expression of their transgenes. The identification and characterization of strong dicot promoter is necessary for the expression of transgenes by native researchers to evaluate their artificial gene. The promoters isolated from viral sources have some limitations. The dicot promoter sequence of ?-tubulin (?-TbSt) was explored and isolated from potato. The ?-TbSt promoter sequence consists of light responsive, hormonal responsive and stress responsive elements. Motifs having responsive elements were identified in ?-TbSt promoter. The ?-TbSt promoter is highly constitutive promoter.
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Lipp, M., R. Schilling, S. Wiest, G. Laux y G. W. Bornkamm. "Target sequences for cis-acting regulation within the dual promoter of the human c-myc gene". Molecular and Cellular Biology 7, n.º 4 (abril de 1987): 1393–400. http://dx.doi.org/10.1128/mcb.7.4.1393-1400.1987.
Resumen
Recombinant plasmids of the human c-myc promoter-leader region and the bacterial chloramphenicol acetyltransferase (cat) gene were constructed. After transfection into different rodent and human cells, the 862-base-pair (bp) PvuII fragment carrying both c-myc promoters and 350 bp of the untranslated leader conferred 1/15 to 1/30 of the CAT activity mediated by the simian virus 40 promoter. The presence of additional sequences upstream of the PvuII fragment had an overall negative effect on c-myc promoter activity detectable by titration analysis with small amounts of transfected plasmid DNA. The analysis of numerous deletion constructs in the c-myc promoter-leader region as well as S1 mapping experiments demonstrated that the high CAT activity depended largely on the presence of the second promoter. By cotransfection of c-myc-cat constructs with plasmids carrying different parts of the c-myc promoter locus, targets for positively acting cellular factors were identified. Two positive regulatory elements were mapped within the 862-bp PvuII fragment. One was localized within the 248-bp PvuII-SmaI fragment -101 to -349 bp upstream of the first cap site and the other within the 142-pb XhoI-NaeI fragment of the first exon, comprising positions -95 to +47 relative to the second cap site. We conclude that the dual promotor of the human c-myc gene represents a strong eucaryotic promotor regulated by cooperation of positively and negatively acting cellular transcription factors.
20
Lipp, M., R. Schilling, S. Wiest, G. Laux y G. W. Bornkamm. "Target sequences for cis-acting regulation within the dual promoter of the human c-myc gene." Molecular and Cellular Biology 7, n.º 4 (abril de 1987): 1393–400. http://dx.doi.org/10.1128/mcb.7.4.1393.
Resumen
Recombinant plasmids of the human c-myc promoter-leader region and the bacterial chloramphenicol acetyltransferase (cat) gene were constructed. After transfection into different rodent and human cells, the 862-base-pair (bp) PvuII fragment carrying both c-myc promoters and 350 bp of the untranslated leader conferred 1/15 to 1/30 of the CAT activity mediated by the simian virus 40 promoter. The presence of additional sequences upstream of the PvuII fragment had an overall negative effect on c-myc promoter activity detectable by titration analysis with small amounts of transfected plasmid DNA. The analysis of numerous deletion constructs in the c-myc promoter-leader region as well as S1 mapping experiments demonstrated that the high CAT activity depended largely on the presence of the second promoter. By cotransfection of c-myc-cat constructs with plasmids carrying different parts of the c-myc promoter locus, targets for positively acting cellular factors were identified. Two positive regulatory elements were mapped within the 862-bp PvuII fragment. One was localized within the 248-bp PvuII-SmaI fragment -101 to -349 bp upstream of the first cap site and the other within the 142-pb XhoI-NaeI fragment of the first exon, comprising positions -95 to +47 relative to the second cap site. We conclude that the dual promotor of the human c-myc gene represents a strong eucaryotic promotor regulated by cooperation of positively and negatively acting cellular transcription factors.
21
Xu, Mengqiong, Sisi Xia, Mei Wang, Xiaolian Liu, Xin Li, Weijie Chen, Yaohao Wang et al. "Enzymatic independent role of sphingosine kinase 2 in regulating the expression of type I interferon during influenza A virus infection". PLOS Pathogens 18, n.º 9 (7 de septiembre de 2022): e1010794. http://dx.doi.org/10.1371/journal.ppat.1010794.
Resumen
Influenza virus has the ability to circumvent host innate immune system through regulating certain host factors for its effective propagation. However, the detailed mechanism is still not fully understood. Here, we report that a host sphingolipid metabolism-related factor, sphingosine kinase 2 (SPHK2), upregulated during influenza A virus (IAV) infection, promotes IAV infection in an enzymatic independent manner. The enhancement of the virus replication is not abolished in the catalytic-incompetent SPHK2 (G212E) overexpressing cells. Intriguingly, the sphingosine-1-phosphate (S1P) related factor HDAC1 also plays a crucial role in SPHK2-mediated IAV infection. We found that SPHK2 cannot facilitate IAV infection in HDAC1 deficient cells. More importantly, SPHK2 overexpression diminishes the IFN-β promoter activity upon IAV infection, resulting in the suppression of type I IFN signaling. Furthermore, ChIP-qPCR assay revealed that SPHK2 interacts with IFN-β promoter through the binding of demethylase TET3, but not with the other promoters regulated by TET3, such as TGF-β1 and IL6 promoters. The specific regulation of SPHK2 on IFN-β promoter through TET3 can in turn recruit HDAC1 to the IFN-β promoter, enhancing the deacetylation of IFN-β promoter, therefore leading to the inhibition of IFN-β transcription. These findings reveal an enzymatic independent mechanism on host SPHK2, which associates with TET3 and HDAC1 to negatively regulate type I IFN expression and thus facilitates IAV propagation.
22
Galvagni, F., M. Lestingi, E. Cartocci y S. Oliviero. "Serum response factor and protein-mediated DNA bending contribute to transcription of the dystrophin muscle-specific promoter." Molecular and Cellular Biology 17, n.º 3 (marzo de 1997): 1731–43. http://dx.doi.org/10.1128/mcb.17.3.1731.
Resumen
The minimal muscle-specific dystrophin promoter contains the consensus sequence CC(A/T)6GG, or the CArG element, which can be found in serum-inducible or muscle-specific promoters. The serum response factor (SRF), which mediates the transcriptional activation of the c-fos gene in response to serum stimulation, can bind to different CArG box elements, suggesting that it could be involved in muscle-constitutive transcription. Here we show that SRF binds to the dystrophin promoter and regulates its muscle-specific transcription. In transient transfections, an altered-binding-specificity SRF mutant restores the muscle-constitutive transcription of a dystrophin promoter with a mutation in its CArG box element. The muscle-constitutive transcription of the dystrophin promoter also requires the sequence GAAACC immediately downstream of the CArG box. This sequence is recognized by a novel DNA bending factor which was named dystrophin promoter-bending factor (DPBF). Mutations of the CArG flanking sequence abolish both DPBF binding and the promoter activity in muscle cells. Its replacement with a p62/ternary complex factor binding site changes the promoter specificity from muscle constitutive to serum responsive. These results show that, on the dystrophin promoter, the transcriptional activation induced by SRF requires the DNA bending induced by DPBF. The bending, next to the CArG box, could promote interactions between SRF and other proteins in the transcriptional complex.
23
Efremova, Larisa N., Svetlana R. Strelnikova, Guzel R. Gazizova, Elena A. Minkina y Roman A. Komakhin. "A Synthetic Strong and Constitutive Promoter Derived from the Stellaria media pro-SmAMP1 and pro-SmAMP2 Promoters for Effective Transgene Expression in Plants". Genes 11, n.º 12 (26 de noviembre de 2020): 1407. http://dx.doi.org/10.3390/genes11121407.
Resumen
Synthetic promoters are vital for genetic engineering-based strategies for crop improvement, but effective methodologies for their creation and systematic testing are lacking. We report here on the comparative analysis of the promoters pro-SmAMP1 and pro-SmAMP2 from Stellaria media ANTIMICROBIAL PEPTIDE1 (AMP1) and ANTIMICROBIAL PEPTIDE2 (AMP2). These promoters are more effective than the well-known Cauliflower mosaic virus 35S promoter. Although these promoters share about 94% identity, the pro-SmAMP1 promoter demonstrated stronger transient expression of a reporter gene in Agrobacterium infiltration of Nicotiana benthamiana leaves, while the pro-SmAMP2 promoter was more effective for the selection of transgenic tobacco (Nicotiana tabacum) cells when driving a selectable marker. Using the cap analysis of gene expression method, we detected no differences in the structure of the transcription start sites for either promoter in transgenic plants. For both promoters, we used fine-scale deletion analysis to identify 160 bp-long sequences that retain the unique properties of each promoter. With the use of chimeric promoters and directed mutagenesis, we demonstrated that the superiority of the pro-SmAMP1 promoter for Agrobacterium-mediated infiltration is caused by the proline-inducible ACTCAT cis-element strictly positioned relative to the TATA box in the core promoter. Surprisingly, the ACTCAT cis-element not only activated but also suppressed the efficiency of the pro-SmAMP1 promoter under proline stress. The absence of the ACTCAT cis-element and CAANNNNATC motif (negative regulator) in the pro-SmAMP2 promoter provided a more constitutive gene expression profile and better selection of transgenic cells on selective medium. We created a new synthetic promoter that enjoys high effectiveness both in transient expression and in selection of transgenic cells. Intact promoters with differing properties and high degrees of sequence identity may thus be used as a basis for the creation of new synthetic promoters for precise and coordinated gene expression.
24
Sorensen, Sven E., Jane M. Barrett, Alex KC Wong y John H. Spencer. "Identification of the in vivo promoters of bacteriophages S13 and ϕX174 and measurement of their relative activities". Biochemistry and Cell Biology 76, n.º 4 (1 de agosto de 1998): 625–36. http://dx.doi.org/10.1139/o98-070.
Resumen
Regions of bacteriophages ϕX174 and S13 that contain putative promoter sequences were amplified by the polymerase chain reaction (PCR) and cloned into the reporter vector pKO-1. Assays of galactokinase activity revealed in vivo promoter activity in those constructs containing the promoter sequences with transcription initiation (+1) sites at nucleotide positions 45, 982, 1823, and 5211. These were identical in location to sequences with in vitro promoter activity and to the three known promoters PA, PB, and PD. P5211 is the location of a new, fourth, promoter. A site with a +1 position at nucleotide 4876, previously shown to initiate RNA synthesis in an in vitro run-off transcription assay, had no in vivo promoter activity. To investigate whether flanking sequences had effects on promoter activity, restriction fragments of ϕX174 and S13 that encompass the in vivo promoters were cloned into the reporter vector pKO-1. The PA and P5211 promoter constructs showed dramatic effects with increases in activity of up to 7 times that shown with the PCR-generated promoter constructs. The ϕX174 PB promoter construct had a 50% decrease in activity compared with the PCR-generated PB clone. While the data showed that in most instances promoter activity is affected by the flanking sequences in which the promoter is embedded, no general pattern correlating flanking sequences and promoter activity could be discerned. Additional evidence that the promoter sequence regions were active in vivo promoters was obtained by S1 nuclease mapping experiments. Initiation of RNA synthesis was shown at positions 45, 982, and 5211.Key words: promoters, phage S13, phage phiX174, in vivo.
25
Mu, X., B. Lee, J. M. Louis y A. R. Kimmel. "Sequence-specific protein interaction with a transcriptional enhancer involved in the autoregulated expression of cAMP receptor 1 in Dictyostelium". Development 125, n.º 18 (15 de septiembre de 1998): 3689–98. http://dx.doi.org/10.1242/dev.125.18.3689.
Resumen
Major stages of Dictyostelium development are regulated by secreted, extracellular cAMP through activation of a serpentine receptor family. During early development, oscillations of extracellular cAMP mobilize cells for aggregation; later, continuous exposure to higher extracellular cAMP concentrations downregulates early gene expression and promotes cytodifferentiation and cell-specific gene expression. The cAMP receptor 1 gene CAR1 has two promoters that are differentially responsive to these extracellular cAMP stimuli. The early CAR1 promoter is induced by nM pulses of cAMP, which in turn are generated by CAR1-dependent activation of adenylyl cyclase (AC). Higher, non-fluctuating concentrations of cAMP will adapt this AC stimulus-response, repress the activated early promoter and induce the dormant late promoter. We now identify a critical element of the pulse-induced CAR1 promoter and a nuclear factor with sequence-specific interaction. Mutation of four nucleotides within the element prevents both in vitro protein binding and in vivo expression of an otherwise fully active early CAR1 promoter and multimerization of the wild-type, but not mutant, sequence will confer cAMP regulation to a quiescent heterologous promoter. These cis and trans elements, thus, constitute a part of the molecular response to the cAMP transmembrane signal cascade that regulates early development of Dictyostelium.
26
Wu, Hai-Young, Jianyou Tan y Ming Fang. "Long-range interaction between two promoters: Activation of the leu-500 promoter by a distant upstream promoter". Cell 82, n.º 3 (agosto de 1995): 445–51. http://dx.doi.org/10.1016/0092-8674(95)90433-6.
27
Carlier, Aurelien L. y S. B. von Bodman. "The rcsA Promoter of Pantoea stewartii subsp. stewartii Features a Low-Level Constitutive Promoter and an EsaR Quorum-Sensing-Regulated Promoter". Journal of Bacteriology 188, n.º 12 (15 de junio de 2006): 4581–84. http://dx.doi.org/10.1128/jb.00211-06.
Resumen
ABSTRACT The upstream region of the Pantoea stewartii rcsA gene features two promoters, one for constitutive basal-level expression and a second autoregulated promoter for induced expression. The EsaR quorum-sensing repressor binds to a site centered between the two promoters, blocking transcription elongation from the regulated promoter under noninducing conditions.
28
Beutel, Tilman, Helmut Knözinger, Aleksei V. Siborov y Vladimir I. Zaikovskii. "Metal–promoter interactions in vanadium oxide promoted Rh/SiO2catalysts". J. Chem. Soc., Faraday Trans. 88, n.º 18 (1992): 2775–82. http://dx.doi.org/10.1039/ft9928802775.
29
Yang, Mary y Laura L. Elnitski. "Diversity of core promoter elements comprising human bidirectional promoters". BMC Genomics 9, Suppl 2 (2008): S3. http://dx.doi.org/10.1186/1471-2164-9-s2-s3.
30
Chowdhary, R., R. A. Ali, W. Albig, D. Doenecke y V. B. Bajic. "Promoter modeling: the case study of mammalian histone promoters". Bioinformatics 21, n.º 11 (15 de marzo de 2005): 2623–28. http://dx.doi.org/10.1093/bioinformatics/bti387.
31
Joshi, B., S. Rastogi, M. Morris, L. M. Carastro, C. Decook, E. Seto y S. P. Chellappan. "Differential regulation of human YY1 and caspase 7 promoters by prohibitin through E2F1 and p53 binding sites". Biochemical Journal 401, n.º 1 (11 de diciembre de 2006): 155–66. http://dx.doi.org/10.1042/bj20060364.
Resumen
Prohibitin is a 30 kDa growth suppressive protein that has pleiotropic functions in the cell. Although prohibitin has been demonstrated to have potent transcriptional regulatory functions, it has also been proposed to facilitate protein folding in the mitochondria and promote cell migration in association with Raf-1. Our previous studies have shown that prohibitin physically interacts with the marked-box domain of E2F family members and represses their transcriptional activity; in contrast, prohibitin could bind to and enhance the transcriptional activity of p53. Here, we show that promoters of human YY1 (Yin and Yang 1) as well as caspase 7 genes are modulated by prohibitin. YY1 promoter activity was reduced upon overexpression of prohibitin, while it was enhanced when prohibitin was depleted by small interfering RNA techniques. The repressive effects of prohibitin on the YY1 promoter were mediated through E2F binding sites, as seen by mutational analysis and chromatin immunoprecipitation assays. Further, depletion of E2F1 prevented prohibitin from repressing the YY1 promoter. In contrast with YY1, prohibitin overexpression led to enhanced levels of caspase 7, whereas depletion of prohibitin reduced it. Interestingly, the caspase 7 promoter was found to have p53-binding sites and prohibitin activated this promoter through p53. These studies show that prohibitin can have diverse effects on the expression of different genes and the activity of various cellular promoters is affected by prohibitin. Further, it appears very likely that prohibitin carries out many of its cellular functions by affecting the transcription of different genes.
32
Wu, Yenan, Lea Kröller, Beiping Miao, Henning Boekhoff, Andrea S. Bauer, Markus W. Büchler, Thilo Hackert, Nathalia A. Giese, Jussi Taipale y Jörg D. Hoheisel. "Promoter Hypermethylation Promotes the Binding of Transcription Factor NFATc1, Triggering Oncogenic Gene Activation in Pancreatic Cancer". Cancers 13, n.º 18 (11 de septiembre de 2021): 4569. http://dx.doi.org/10.3390/cancers13184569.
Resumen
Studies have indicated that some genes involved in carcinogenesis are highly methylated in their promoter regions but nevertheless strongly transcribed. It has been proposed that transcription factors could bind specifically to methylated promoters and trigger transcription. We looked at this rather comprehensively for pancreatic ductal adenocarcinoma (PDAC) and studied some cases in more detail. Some 2% of regulated genes in PDAC exhibited higher transcription coupled to promoter hypermethylation in comparison to healthy tissue. Screening 661 transcription factors, several were found to bind specifically to methylated promoters, in particular molecules of the NFAT family. One of them—NFATc1—was substantially more strongly expressed in PDAC than control tissue and exhibited a strong oncogenic role. Functional studies combined with computational analyses allowed determining affected genes. A prominent one was gene ALDH1A3, which accelerates PDAC metastasis and correlates with a bad prognosis. Further studies confirmed the direct up-regulation of ALDH1A3 transcription by NFATc1 promoter binding in a methylation-dependent process, providing insights into the oncogenic role of transcription activation in PDAC that is promoted by DNA methylation.
33
Fan, Xinyu y Feng Yang. "Strategic promotion, reputation, and responsiveness in bureaucratic hierarchies". Journal of Theoretical Politics 31, n.º 3 (5 de junio de 2019): 286–307. http://dx.doi.org/10.1177/0951629819850638.
Resumen
While existing studies usually model promotion as a bilateral interaction between promoter and promotee, it is not uncommon that the promoter is under the influence of a third party. For instance, authoritarian rulers may consider how their interactions with local agents change the way that citizens view them. Similarly, a mid-tier officer in a bureaucratic hierarchy often concerns herself with her image in the eyes of her superior when managing her subordinates. In this paper, we construct a game-theoretic model to investigate promotion strategies when promoters have reputation concerns. We show that promoters can use promotion as a signaling tool, where she can deliberately postpone promoting the subordinate to enhance her own reputation. Furthermore, the promoter has extra incentives to shirk, knowing that she can manipulate promotion in the future. Thus, strategic promotions decrease government responsiveness. Counter-intuitively, such a decrease is more severe when intra-bureaucracy information is more transparent. In other words, transparency may do more harm than good. We conduct a case study of the Chinese bureaucracy and provide supportive evidence.
34
Choi, Jae Young, Soo-Jin Kwon, Jong Yul Roh, Tae-Jin Yang, Ming Shun Li, Beom-Seok Park, Yonggyun Kim, Soo-Dong Woo, Byung Rae Jin y Yeon Ho Je. "Analysis of promoter activity of selected Cotesia plutellae bracovirus genes". Journal of General Virology 90, n.º 5 (1 de mayo de 2009): 1262–69. http://dx.doi.org/10.1099/vir.0.009472-0.
Resumen
In a previous study, we cloned 27 discrete genome segments of Cotesia plutellae bracovirus (CpBV) and provided the complete nucleotide sequences and annotation. Seven putative coding regions were predicted from one of the largest segments, CpBV-S30. The activity of promoters associated with six predicted ORFs from this segment were investigated using both transient and baculovirus expression assays with enhanced green fluorescent protein as a reporter gene. CpBV promoters showed activity earlier than the polyhedrin promoter and the activity of some of these promoters was superior to that of the Autographa californica multiple nucleopolyhedrovirus (AcMNPV) ie-1 promoter in the baculovirus expression assays. The promoter of ORF3004 showed the highest level of activity in insect cells, exhibiting 24 % of the activity obtained with the AcMNPV polyhedrin promoter in Sf9 cells. In Spodoptera exigua larvae, the ORF3006 promoter showed the highest activity, with about 35 % of the activity measured with the polyhedrin promoter. In addition, analysis of the ORF3006 promoter revealed that the region between −382 and −422 from the translation start point was critical for activity of this promoter. These results suggest that the CpBV-S30 promoters characterized here could be useful tools in a variety of biotechnological applications, such as gene expression analyses and insecticide development.
35
Han, Yoo-Jeong y Primal de Lanerolle. "Naturally Extended CT · AG Repeats Increase H-DNA Structures and Promoter Activity in the Smooth Muscle Myosin Light Chain Kinase Gene". Molecular and Cellular Biology 28, n.º 2 (8 de noviembre de 2007): 863–72. http://dx.doi.org/10.1128/mcb.00960-07.
Resumen
ABSTRACT Naturally occurring repeat sequences capable of adopting H-DNA structures are abundant in promoters of disease-related genes. In support of this, we found (CT)22 · (AG)22 repeats in the promoter of smooth muscle myosin light chain kinase (smMLCK), a key regulator of vascular smooth muscle function. We also found an insertion mutation that adds another six pairs of CT · AG repeats and increases smMLCK promoter activity in spontaneously hypertensive rats (SHR). Therefore, we used the smMLCK promoters from normotensive and hypertensive rats as a model system to determine how CT · AG repeats form H-DNA, an intramolecular triplex, and regulate promoter activity. High-resolution mapping with a chemical probe selective for H-DNA showed that the CT · AG repeats adopt H-DNA structures at a neutral pH. Importantly, the SHR promoter forms longer H-DNA structures than the promoter from normotensive rats. Reconstituting nucleosomes on the promoters, in vitro, showed no difference in nucleosome positioning between the two promoters. However, chromatin immunoprecipitation analyses revealed that histone acetylations are greater in the hypertensive promoter. Thus, our findings suggest that the extended CT · AG repeats in the SHR promoter increase H-DNA structures, histone modifications, and promoter activity of the smMLCK, perhaps contributing to vascular disorders in hypertension.
36
Makara, P. "Editorial. Can we promote equity when we promoter health?" Health Promotion International 12, n.º 2 (1 de junio de 1997): 97–98. http://dx.doi.org/10.1093/heapro/12.2.97.
37
Charoenpanich, Jittima, Akio Tani, Naoko Moriwaki, Kazuhide Kimbara y Fusako Kawai. "Dual regulation of a polyethylene glycol degradative operon by AraC-type and GalR-type regulators in Sphingopyxis macrogoltabida strain 103". Microbiology 152, n.º 10 (1 de octubre de 2006): 3025–34. http://dx.doi.org/10.1099/mic.0.29127-0.
Resumen
The genes for polyethylene glycol (PEG) catabolism (pegB, C, D, A and E) in Sphingopyxis macrogoltabida strain 103 were shown to form a PEG-inducible operon. The pegR gene, encoding an AraC-type regulator in the downstream area of the operon, is transcribed in the reverse direction. The transcription start sites of the operon were mapped, and three putative σ 70-type promoter sites were identified in the pegB, pegA and pegR promoters. A promoter activity assay showed that the pegB promoter was induced by PEG and oligomeric ethylene glycols, whereas the pegA and pegR promoters were induced by PEG. Deletion analysis of the pegB promoter indicated that the region containing the activator-binding motif of an AraC/XylS-type regulator was required for transcription of the pegBCDAE operon. Gel retardation assays demonstrated the specific binding of PegR to the pegB promoter. Transcriptional fusion studies of pegR with pegA and pegB promoters suggested that PegR regulates the expression of the pegBCDAE operon positively through its binding to the pegB promoter, but PegR does not bind to the pegA promoter. Two specific binding proteins for the pegA promoter were purified and identified as a GalR-type regulator and an H2A histone fragment (histone-like protein, HU). The binding motif of a GalR/LacI-type regulator was found in the pegA and pegR promoters. These results suggested the dual regulation of the pegBCDAE operon through the pegB promoter by an AraC-type regulator, PegR (PEG-independent), and through the pegA and pegR promoters by a GalR/LacI-type regulator together with HU (PEG-dependent).
38
Pan, Xuewei, Mi Tang, Jiajia You, Yanan Hao, Xian Zhang, Taowei Yang y Zhiming Rao. "A Novel Method to Screen Strong Constitutive Promoters in Escherichia coli and Serratia marcescens for Industrial Applications". Biology 12, n.º 1 (30 de diciembre de 2022): 71. http://dx.doi.org/10.3390/biology12010071.
Resumen
Promoters serve as the switch of gene transcription, playing an important role in regulating gene expression and metabolites production. However, the approach to screening strong constitutive promoters in microorganisms is still limited. In this study, a novel method was designed to identify strong constitutive promoters in E. coli and S. marcescens based on random genomic interruption and fluorescence-activated cell sorting (FACS) technology. First, genomes of E. coli, Bacillus subtilis, and Corynebacterium glutamicum were randomly interrupted and inserted into the upstream of reporter gene gfp to construct three promoter libraries, and a potential strong constitutive promoter (PBS) suitable for E. coli was screened via FACS technology. Second, the core promoter sequence (PBS76) of the screened promoter was identified by sequence truncation. Third, a promoter library of PBS76 was constructed by installing degenerate bases via chemical synthesis for further improving its strength, and the intensity of the produced promoter PBS76-100 was 59.56 times higher than that of the promoter PBBa_J23118. Subsequently, promoters PBBa_J23118, PBS76, PBS76-50, PBS76-75, PBS76-85, and PBS76-100 with different strengths were applied to enhance the metabolic flux of L-valine synthesis, and the L-valine yield was significantly improved. Finally, a strong constitutive promoter suitable for S. marcescens was screened by a similar method and applied to enhance prodigiosin production by 34.81%. Taken together, the construction of a promoter library based on random genomic interruption was effective to screen the strong constitutive promoters for fine-tuning gene expression and reprogramming metabolic flux in various microorganisms.
39
Sun, Wu-Sheng, Hyeon Yang, Jin Gu No, Haesun Lee, Nahyun Lee, Minguk Lee, Man-Jong Kang y Keon Bong Oh. "Select Porcine Elongation Factor 1α Sequences Mediate Stable High-Level and Upregulated Expression of Heterologous Genes in Porcine Cells in Response to Primate Serum". Genes 12, n.º 7 (7 de julio de 2021): 1046. http://dx.doi.org/10.3390/genes12071046.
Resumen
Genetically engineered (GE) pigs with various combinations of genetic profiles have been developed using heterologous promoters. This study aimed to identify autologous promoters for high and ubiquitous expression of xenotransplantation relevant genes in GE pigs. A 1.4 kb upstream regulatory sequence of porcine elongation factor 1α (pEF1α) gene was selected and isolated for use as a promoter. Activity of the pEF1α promoter was subsequently compared with that of the cytomegalovirus (CMV) promoter, CMV enhancer/chicken β-actin (CAG) promoter, and human EF1α (hEF1α) promoter in different types of pig-derived cells. Comparative analysis of luciferase and mutant human leukocyte antigen class E-F2A-β-2 microglobulin (HLA-E) expression driven by pEF1α, CMV, CAG, and hEF1α promoters revealed the pEF1α promoter mediated comparable expression levels with those of the CAG promoter in porcine ear skin fibroblasts (PEFs) and porcine kidney-15 (PK-15) cells, but lower than those of the CAG promoter in porcine aortic endothelial cells (PAECs). The pEF1α promoter provided long-term stable HLA-E expression in PEFs, but the CAG promoter failed to sustain those levels of expression. For xenogeneic serum-induced cytotoxicity assays, the cells were cultured for several hours in growth medium supplemented with primate serum. Notably, the pEF1α promoter induced significant increases in luciferase and HLA-E expression in response to primate serum in PAECs compared with those driven by the CAG promoter, suggesting the pEF1α promoter could regulate temporal expression of heterologous genes under xenogeneic-cytotoxic conditions. These results suggest the pEF1α promoter may be valuable for development of GE pigs spatiotemporally and stably expressing immunomodulatory genes for xenotransplantation.
40
MacLellan, Shawn R., Allyson M. MacLean y Turlough M. Finan. "Promoter prediction in the rhizobia". Microbiology 152, n.º 6 (1 de junio de 2006): 1751–63. http://dx.doi.org/10.1099/mic.0.28743-0.
Resumen
The ability to recognize and predict non-σ 54 promoters in the alphaproteobacteria is not well developed. In this study, 25 experimentally verified Sinorhizobium meliloti promoter sequences were compiled and used to predict the location of other related promoters in the S. meliloti genome. Fourteen candidate predictions were targeted for verification and of these at least 12 proved to be genuine promoters. As a result, the experimental identification of 12 novel promoters linked to genes rpoD, topA, rpmJ, trpS, ropB1, metC, rpsT, secE, trkH and three tRNA genes is reported. In all, 99 predicted and verified promoters are reported, including those linked with 13 tRNA genes, eight ribosomal protein genes and a number of other physiologically important or essential genes. On the basis of sequence conservation and a mutational analysis of promoter activity, the −35 and −10 consensus for these promoters is 5-CTTGAC-N17-CTATAT. This promoter structure, which seems to be widely conserved amongst several other genera in the alphaproteobacteria, shares significant similarity with, but is skewed by a 1 nt step from, the canonical Escherichia coli σ 70 promoter. Perhaps this difference is responsible for the observation that S. meliloti promoters are often poorly expressed in E. coli. In this regard, expression data from plasmid-borne gfp-reporter fusions to eight of the S. meliloti promoters verified in this work revealed that while these promoters were very active in S. meliloti and Agrobacterium tumefaciens only very low, near-background activity was detected in E. coli.
41
Sebastian, Siby, Kazuto Takayama, Makio Shozu y Serdar E. Bulun. "Cloning and Characterization of a Novel Endothelial Promoter of the Human CYP19 (Aromatase P450) Gene that Is Up-Regulated in Breast Cancer Tissue". Molecular Endocrinology 16, n.º 10 (1 de octubre de 2002): 2243–54. http://dx.doi.org/10.1210/me.2002-0123.
Resumen
Abstract Intratumoral expression of aromatase P450 (P450arom) promotes the growth of breast tumors via increased local estrogen concentration. We cloned a novel 101-bp untranslated first exon (I.7) that comprises the 5′-end of 29–54% of P450arom transcripts isolated from breast cancer tissues (n = 7). The levels of P450arom transcripts with exon I.7 were significantly increased in breast tumor tissues and adipose tissue adjacent to tumors. We identified a promoter immediately upstream of exon I.7 and mapped this to about 36 kb upstream of ATG translation start site of the CYP19 (aromatase cytochrome P450) gene. Sequence analysis of I.7 revealed a TATA-less promoter containing an initiator, two consensus GATA sites, and cis-regulatory elements found in megakaryocytes and endothelial type promoters. Luciferase activity directed by the promoter I.7 sequence (−299/+81 bp) was 4-fold greater than a minimum length promoter sequence (−35/+81 bp) in human microvascular endothelial cells (HMEC-1), but only 2-fold greater in MCF-7 breast malignant epithelial cells. There was no promoter activity in primary breast adipose fibroblasts. Site-directed mutations demonstrated that maximal basal promoter activity required two GATA motifs at −146/−141 bp and −196/−191 bp. Gel shift and deoxyribonuclease I footprinting assays demonstrated the binding of GATA-2 transcription factor but not GATA-1 to the −196/−191-bp region. Overexpression of GATA-2 in HMEC-1 cells increases promoter I.7 activity by 5-fold. In conclusion, promoter I.7 is a GATA-2-regulated endothelial promoter of the human CYP19 gene and may increase estrogen biosynthesis in vascular endothelial cells of breast cancer. The activity of this promoter may also be important for intracrine and paracrine effects of estrogen on blood vessels.
42
Tseng, Ching Wen, Shuping Zhang y George C. Stewart. "Accessory Gene Regulator Control of Staphyloccoccal Enterotoxin D Gene Expression". Journal of Bacteriology 186, n.º 6 (15 de marzo de 2004): 1793–801. http://dx.doi.org/10.1128/jb.186.6.1793-1801.2004.
Resumen
ABSTRACT The quorum-sensing system of Staphylococcus aureus, the accessory gene regulator (Agr) system, is responsible for increased transcription of certain exoprotein genes and decreased transcription of certain cell wall-associated proteins during the postexponential phase of growth. This regulation is important for virulence, as evidenced by a reduction in virulence associated with a loss of the Agr system. The enterotoxin D (sed) determinant is upregulated by the Agr system. To define the Agr-regulated cis element(s) within the sed promoter region, we utilized promoters not regulated by Agr to create hybrid promoters. Hybrid promoters were created by using sed sequences combined with the enterotoxin A (sea) promoter or the S. aureus lac operon promoter sequences. The results obtained indicated that the Agr control element of the sed promoter resides within the −35 promoter element and at the Pribnow box to the +1 site of the promoter. At these positions of the sed promoter, a directly repeated 6-bp sequence was found. This repeat is important for overall promoter activity, and maximal regulation of the promoter activity requires both repeat elements. Furthermore, Agr control of sed promoter activity was found to be dependent upon the presence of a functional Rot protein. Therefore, the postexponential increase in sed transcription results from the Agr-mediated reduction in Rot activity rather than as a direct effect of the Agr system.
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Yan, Bing-xue y Jin-xia Ma. "Promoter-associated RNAs and promoter-targeted RNAs". Cellular and Molecular Life Sciences 69, n.º 17 (14 de marzo de 2012): 2833–42. http://dx.doi.org/10.1007/s00018-012-0953-1.
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Wu, Limin, Aliaa El-Mezawy y Saleh Shah. "Isolation and evaluation of three novel native promoters in Brassica napus". Botany 91, n.º 6 (junio de 2013): 414–19. http://dx.doi.org/10.1139/cjb-2012-0245.
Resumen
To provide effective and specific native promoters for canola (Brassica napus L.) genetic modification, three promoters were isolated by genome walking from this species. These three promoters were fused to the uidA reporter gene (GUS) and were independently used to generate populations of transgenic canola plants. Plants transformed with BnPGPro-GUS (B. napus putative germin promoter) exhibited GUS activity in all the tissues tested at a level comparable to those transformed with CaMV35 S promoter. This indicates that BnPGPro may serve as a native constitutive promoter for canola. The other two promoters, BnPro3-GUS and BnPro5-GUS (B. napus, promoter 3 and 5), exhibited GUS activity in various tissues. None of these two promoters expressed in embryo, however. These novel Brassica native promoters can be used to modify canola genes for various purposes.
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Mans, Ruud M. W. y Dagmar Knebel-Mörsdorf. "In Vitro Transcription of pe38/Polyhedrin Hybrid Promoters Reveals Sequences Essential for Recognition by the Baculovirus-Induced RNA Polymerase and for the Strength of Very Late Viral Promoters". Journal of Virology 72, n.º 4 (1 de abril de 1998): 2991–98. http://dx.doi.org/10.1128/jvi.72.4.2991-2998.1998.
Resumen
ABSTRACT In vitro transcription was used to analyze the promoter specificity of the α-amanitin-resistant RNA polymerase that is induced late during infection of Autographa californica multicapsid nuclear polyhedrosis virus. By modifying the preparation of crude nuclear extracts, we have established an assay that permits differentiation between weak late and strong very late viral promoters. The virus-induced RNA polymerase initiates at a TAAG sequence motif in both late and very late promoters. Based on the sensitivity of our in vitro transcription system, we have investigated the sequences responsible for a functional TAAG motif and their putative role with respect to the strength of very late promoters. By constructing hybrid promoters between the early pe38 and the very late polyhedrin promoters, we demonstrated that the replacement of 7 nucleotides upstream of the nonfunctional TAAG sequences in the pe38 promoter with the corresponding sequences of the polyhedrin promoter was sufficient for recognition by the virus-induced RNA polymerase. The strength of the very late polyhedrin promoter was established after replacing the 5′ untranslated sequences of the pe38 promoter by those of the polyhedrin promoter in addition to the 7 nucleotides upstream of the TAAG motif.
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He, Kevin, S. M. Ali Hosseini Rad, Aarati Poudel y Alexander Donald McLellan. "Compact Bidirectional Promoters for Dual-Gene Expression in a Sleeping Beauty Transposon". International Journal of Molecular Sciences 21, n.º 23 (4 de diciembre de 2020): 9256. http://dx.doi.org/10.3390/ijms21239256.
Resumen
Promoter choice is an essential consideration for transgene expression in gene therapy. The expression of multiple genes requires ribosomal entry or skip sites, or the use of multiple promoters. Promoter systems comprised of two separate, divergent promoters may significantly increase the size of genetic cassettes intended for use in gene therapy. However, an alternative approach is to use a single, compact, bidirectional promoter. We identified strong and stable bidirectional activity of the RPBSA synthetic promoter comprised of a fragment of the human Rpl13a promoter, together with additional intron/exon structures. The Rpl13a-based promoter drove long-term bidirectional activity of fluorescent proteins. Similar results were obtained for the EF1-α and LMP2/TAP1 promoters. However, in a lentiviral vector, the divergent bidirectional systems failed to produce sufficient titres to translate into an expression system for dual chimeric antigen receptor (CAR) expression. Although bidirectional promoters show excellent applicability to drive short RNA in Sleeping Beauty transposon systems, their possible use in the lentiviral applications requiring longer and more complex RNA, such as dual-CAR cassettes, is limited.
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Ansarikaleibari, Aida. "Architecture of Bacterial Promoters; The case of the Escherichia coli ogt Promoter". Journal of Molecular Biology Research 6, n.º 1 (22 de noviembre de 2016): 111. http://dx.doi.org/10.5539/jmbr.v6n1p111.
Resumen
<p class="1Body">All bacteria utilize RNA polymerase enzyme and transcription activator proteins to regulate gene expression in response to internal or external stress. Some bacterial promoters are regulated with only one transcription factor whilst two or more transcription activators regulate some other promoters. NarL is a transcription activator protein that activates the <em>E. coli yeaR</em> and <em>ogt</em> promoters in response to nitrate and nitrite induction in absence of oxygen. In the present study we have studied <em>ogt1052</em> promoter, which is a derivative of <em>ogt</em> promoter containing only one NarL binding site very close to -35 element. Therefore, it is considered as class II activator dependent promoter just as <em>yeaR</em> promoter. A molecular structure of <em>ogt1052</em> promoter was proposed which suggests that NarL binding site is located in opposite face of DNA that contains Alpha-CTD and sigma domain 4 of RNA polymerase enzyme required for promoter recognition. The aim of the present study was to study and test the suggested molecular model by creating point mutations at -35 element and deletion of one base pair in spacer region, to test whether sigma domain 4 is necessary to bind -35 hexamer in order to start transcription initiation, and to test whether NarL activates the promoter by interaction with Alpha-CTD in the opposite face of the DNA. Based on the result achieved, <em>ogt1052</em> promoter is a class I promoter “dressed” like a class II promoter.</p>
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Haniyya, Haniyya, Dini Achnafani, Maria Ulfah, Niknik Nurhayati y Is Helianti. "The utilization of auto-inducible Plyb promoter and media optimation for cell density-dependent expression of recombinant xylanase in Bacillus subtilis DB104". Microbiology Indonesia 14, n.º 1 (1 de julio de 2020): 2. http://dx.doi.org/10.5454/mi.14.1.2.
Resumen
Strong promoters are one of the fundamental aspects to increase the level of gene expression, and one of approach to improve the recombinant enzyme productivity so that the efficiency of production cost for enzyme production in industrial scale can be reached. Here we assessed the application of a cell density-dependent promoter and media optimation to promote cell growth and protein expression of Bacillus subtilis without excess usage of inducers. An auto-inducible Pylb promoter that is potential to provide inducer-free enzyme production was cloned and introduced into xylanase recombinant system in B. subtilis DB104 by PCR cloning and protoplast transformation. A 200 bp target gene was successfully inserted in between xynCM1 ORF -coding for B. halodurans CM1 xylanase- and its native promoter sequence at the upstream region. The disruption of the native promoter was intended to replace the native promoter with Pylb. Recombinant xylanase gene under Pylb was successfully expressed in B. subtilis DB104 and the enzyme was produced at stationary phase. Different media with various concentrations of glucose and nitrogen were used to optimize recombinant xylanase expression. It achieved a higher level of xylanase expression compared to wild-type and recombinant xylanase with native promoter B. subtilis in media containing a 2-fold recipe of LB media thus leads to increase cell density and xylanase expression (81.461 U mL-1).
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Valladares, Ana, Alicia M. Muro-Pastor, Antonia Herrero y Enrique Flores. "The NtcA-Dependent P1 Promoter Is Utilized for glnA Expression in N2-Fixing Heterocysts of Anabaena sp. Strain PCC 7120". Journal of Bacteriology 186, n.º 21 (1 de noviembre de 2004): 7337–43. http://dx.doi.org/10.1128/jb.186.21.7337-7343.2004.
Resumen
ABSTRACT Expression of the glnA gene encoding glutamine synthetase, a key enzyme in nitrogen metabolism, is subject to a variety of regulatory mechanisms in different organisms. In the filamentous, N2-fixing cyanobacterium Anabaena sp. strain PCC 7120, glnA is expressed from multiple promoters that generate several transcripts whose abundance is influenced by NtcA, the transcription factor exerting global nitrogen control in cyanobacteria. Whereas RNAI originates from a canonical NtcA-dependent promoter (P1) and RNAII originates from a σ70-type promoter (P2), RNAIV is influenced by NtcA but the corresponding promoter (P3) does not have the structure of NtcA-activated promoters. Using RNA isolated from Anabaena filaments grown under different nitrogen regimens, we observed, in addition to these transcripts, RNAV, which has previously been detected only in in vitro transcription assays and should originate from P4. However, in heterocysts, which are differentiated cells specialized in N2 fixation, RNAI was the almost exclusive glnA transcript. Analysis of P glnA ::lacZ fusions containing different fragments of the glnA upstream region confirmed that fragments carrying P1, P2, or P3 and P4 have the ability to promote transcription. Mutation of the NtcA-binding site in P1 eliminated P1-directed transcription and allowed increased use of P2. The NtcA-binding site in the P1 promoter and binding of NtcA to this site appear to be key factors in determining glnA gene expression in vegetative cells and heterocysts.
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Koev, Gennadiy y W. Allen Miller. "A Positive-Strand RNA Virus with Three Very Different Subgenomic RNA Promoters". Journal of Virology 74, n.º 13 (1 de julio de 2000): 5988–96. http://dx.doi.org/10.1128/jvi.74.13.5988-5996.2000.
Resumen
ABSTRACT Numerous RNA viruses generate subgenomic mRNAs (sgRNAs) for expression of their 3′-proximal genes. A major step in control of viral gene expression is the regulation of sgRNA synthesis by specific promoter elements. We used barley yellow dwarf virus (BYDV) as a model system to study transcriptional control in a virus with multiple sgRNAs. BYDV generates three sgRNAs during infection. The sgRNA1 promoter has been mapped previously to a 98-nucleotide (nt) region which forms two stem-loop structures. It was determined that sgRNA1 is not required for BYDV RNA replication in oat protoplasts. In this study, we show that neither sgRNA2 nor sgRNA3 is required for BYDV RNA replication. The promoters for sgRNA2 and sgRNA3 synthesis were mapped by using deletion mutagenesis. The minimal sgRNA2 promoter is approximately 143 nt long (nt 4810 to 4952) and is located immediately downstream of the putative sgRNA2 start site (nt 4809). The minimal sgRNA3 core promoter is 44 nt long (nt 5345 to 5388), with most of the sequence located downstream of sgRNA3 start site (nt 5348). For both promoters, additional sequences upstream of the start site enhanced sgRNA promoter activity. These promoters contrast to the sgRNA1 promoter, in which almost all of the promoter is located upstream of the transcription initiation site. Comparison of RNA sequences and computer-predicted secondary structures revealed little or no homology between the three sgRNA promoter elements. Thus, a small RNA virus with multiple sgRNAs can have very different subgenomic promoters, which implies a complex system for promoter recognition and regulation of subgenomic RNA synthesis.