Artigos de revistas: "RNU2"

1

Li, Zengji, Adong Yu e Alan M. Weiner. "Adenovirus Type 12-Induced Fragility of the Human RNU2 Locus Requires p53 Function". Journal of Virology 72, n.º 5 (1 de maio de 1998): 4183–91. http://dx.doi.org/10.1128/jvi.72.5.4183-4191.1998.

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ABSTRACT Adenovirus type 12 (Ad12) infection of human cells induces four chromosomal fragile sites corresponding to the U1 small nuclear RNA (snRNA) genes (the RNU1 locus), the U2 snRNA genes (RNU2), the U1 snRNA pseudogenes (PSU1), and the 5S rRNA genes (RN5S). Ad12-induced fragility of the RNU2 locus requires U2 snRNA transcriptional regulatory elements and viral early functions but not viral replication or integration, or chromosomal sequences flanking the RNU2 locus. We now show that Ad12 cannot induce the RNU1, RNU2, or PSU1fragile sites in Saos-2 cells lacking the p53 and retinoblastoma (Rb) proteins but that viral induction of fragility is rescued in these cells when the expression of wild-type p53 or selected hot-spot mutants (i.e., V143A, R175H, R248W, and R273H) is restored by transient expression or stable retroviral transduction. We also observed weak constitutive fragility of theRNU1 and RNU2 loci in cells belonging to xeroderma pigmentosum complementation groups B and D (XPB and XPD) which are partially defective in the ERCC2 (XPD) and ERCC3 (XPB) helicase activities shared between the repairosome and the RNA polymerase II basal transcription factor TFIIH. We propose a model for Ad12-induced chromosome fragility in which interaction of p53 with the Ad12 E1B 55-kDa transforming protein (and possibly E4orf6) induces a p53 gain of function which ultimately perturbs the RNA polymerase II basal transcription apparatus. The p53 gain of function could interfere with chromatin condensation either by blocking mitotic shutdown of U1 and U2 snRNA transcription or by phenocopying global or local DNA damage. Specific fragilization of the RNU1, RNU2, and PSU1 loci could reflect the unusually high local concentration of strong transcription units or the specialized nature of the U1 and U2 snRNA transcription apparatus.

2

Kuhlmann, Jan Dominik, Alexander Baraniskin, Stephan A. Hahn, Frank Mosel, Maren Bredemeier, Pauline Wimberger, Rainer Kimmig e Sabine Kasimir-Bauer. "Circulating U2 Small Nuclear RNA Fragments as a Novel Diagnostic Tool for Patients with Epithelial Ovarian Cancer". Clinical Chemistry 60, n.º 1 (1 de janeiro de 2014): 206–13. http://dx.doi.org/10.1373/clinchem.2013.213066.

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Abstract BACKGROUND Ovarian cancer is the leading cause of death among malignancies in women. Despite advances in treatment, >50% of patients relapse. For disease monitoring, the identification of a blood-based biomarker would be of prime interest. In this regard, noncoding RNAs, such as microRNA (miRNA) or small nuclear RNA (snRNA), have been suggested as biomarkers for noninvasive cancer diagnosis. In the present study, we sought to identify differentially expressed miRNA/snRNA in sera of ovarian cancer patients and investigate their potential to aid in therapy monitoring. METHODS miRNA/snRNA abundance was investigated in serum (n = 10) by microarray analysis and validated in an extended serum set (n = 119) by reverse-transcription quantitative PCR. RESULTS Abundance of U2-1 snRNA fragment (RNU2-1f) was significantly increased in sera of ovarian cancer patients (P < 0.0001) and paralleled International Federation of Gynecology and Obstetrics stage as well as residual tumor burden after surgery (P < 0.0001 and P = 0.011, respectively). Moreover, for patients with suboptimal debulking, preoperative RNU2-1f concentration was associated with radiographic response after chemotherapy and with platinum resistance (P = 0.0088 and P = 0.0015, respectively). Interestingly, according to the RNU2-1f abundance dynamics, persistent RNU2-1f positivity before surgery and after chemotherapy identified a subgroup of patients with high risk of recurrence and poor prognosis. CONCLUSIONS This is the first report to suggest that a circulating snRNA can serve as an auxiliary diagnostic tool for monitoring tumor dynamics in ovarian cancer. Our results provide a rationale to further investigate whether this high-risk patient group may benefit from additional therapies that are directly applied after chemotherapy.

3

Yu, A. "Metaphase fragility of the human RNU1 and RNU2 loci is induced by actinomycin D through a p53-dependent pathway". Human Molecular Genetics 7, n.º 4 (1 de abril de 1998): 609–17. http://dx.doi.org/10.1093/hmg/7.4.609.

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4

Lai, Yu‐Chang, Yu‐Ting Lai, Md Mahfuzur Rahman, Hui‐Wen Chen, Al Asmaul Husna, Takuro Fujikawa, Takaaki Ando et al. "Bovine milk transcriptome analysis reveals microRNAs and RNU2 involved in mastitis". FEBS Journal 287, n.º 9 (15 de novembro de 2019): 1899–918. http://dx.doi.org/10.1111/febs.15114.

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5

del Río-Moreno, Mercedes, Raúl M. Luque, Oriol A. Rangel-Zúñiga, Emilia Alors-Pérez, Juan F. Alcalá-Diaz, Irene Roncero-Ramos, Antonio Camargo, Manuel D. Gahete, José López-Miranda e Justo P. Castaño. "Dietary Intervention Modulates the Expression of Splicing Machinery in Cardiovascular Patients at High Risk of Type 2 Diabetes Development: From the CORDIOPREV Study". Nutrients 12, n.º 11 (17 de novembro de 2020): 3528. http://dx.doi.org/10.3390/nu12113528.

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Type-2 diabetes mellitus (T2DM) has become a major health problem worldwide. T2DM risk can be reduced with healthy dietary interventions, but the precise molecular underpinnings behind this association are still incompletely understood. We recently discovered that the expression profile of the splicing machinery is associated with the risk of T2DM development. Thus, the aim of this work was to evaluate the influence of 3-year dietary intervention in the expression pattern of the splicing machinery components in peripheral blood mononuclear cells (PBMCs) from patients within the CORDIOPREV study. Expression of splicing machinery components was determined in PBMCs, at baseline and after 3 years of follow-up, from all patients who developed T2DM (Incident-T2DM, n = 107) and 108 randomly selected non-T2DM subjects, who were randomly enrolled in two healthy dietary patterns (Mediterranean or low-fat diets). Dietary intervention modulated the expression of key splicing machinery components (i.e., up-regulation of SPFQ/RMB45/RNU6, etc., down-regulation of RNU2/SRSF6) after three years, independently of the type of healthy diet. Some of these changes (SPFQ/RMB45/SRSF6) were associated with key clinical features and were differentially induced in Incident-T2DM patients and non-T2DM subjects. This study reveals that splicing machinery can be modulated by long-term dietary intervention, and could become a valuable tool to screen the progression of T2DM.

6

Liu, Xudong, e David F. Barker. "Evidence for Effective Suppression of Recombination in the Chromosome 17q21 Segment Spanning RNU2–BRCA1". American Journal of Human Genetics 64, n.º 5 (maio de 1999): 1427–39. http://dx.doi.org/10.1086/302358.

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7

Tessereau, Chloé, Monique Buisson, Nastasia Monnet, Marine Imbert, Laure Barjhoux, Caroline Schluth-Bolard, Damien Sanlaville et al. "Direct Visualization of the Highly Polymorphic RNU2 Locus in Proximity to the BRCA1 Gene". PLoS ONE 8, n.º 10 (11 de outubro de 2013): e76054. http://dx.doi.org/10.1371/journal.pone.0076054.

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8

Bailey, A. D., Z. Li, T. Pavelitz e A. M. Weiner. "Adenovirus type 12-induced fragility of the human RNU2 locus requires U2 small nuclear RNA transcriptional regulatory elements." Molecular and Cellular Biology 15, n.º 11 (novembro de 1995): 6246–55. http://dx.doi.org/10.1128/mcb.15.11.6246.

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Infection of human cells with oncogenic adenovirus type 12 (Ad12) induces four specific chromosome fragile sites. Remarkably, three of these sites appear to colocalize with tandem arrays of genes encoding small, abundant, ubiquitously expressed structural RNAs--the RNU1 locus encoding U1 small nuclear RNA (snRNA), the RNU2 locus encoding U2 snRNA, and the RN5S locus encoding 5S rRNA. Recently, an artificial tandem array of the natural 5.8-kb U2 repeat unit has been shown to generate a new Ad12-inducible fragile site (Y.-P. Li, R. Tomanin, J. R. Smiley, and S. Bacchetti, Mol. Cell. Biol. 13:6064-6070, 1993), demonstrating that the U2 repeat unit alone is sufficient for virally induced fragility. To identify elements within the U2 repeat unit that are required for virally induced fragility, we generated cell lines containing artificial tandem arrays of the entire 5.8-kb repeat unit, an 834-bp fragment spanning the U2 gene alone, or the same 834-bp fragment from which key U2 transcriptional regulatory elements had been deleted. The U2 snRNA coding regions within each artificial array were marked by an innocuous single base change (U to C at position 87) so that the relative expression of supernumerary and endogenous U2 genes could be monitored by a primer extension assay. We find that artificial arrays of both the 5.8- and the 0.8-kb U2 repeat units are fragile but that arrays lacking either the distal sequence element or both the distal and the proximal sequence elements of the promoter are not. Surprisingly, variations in repeat copy number and/or transcriptional activity of the artificial arrays do not appear to correlate with the degree of Ad12-inducible fragility. We conclude that U2 transcriptional regulatory elements are required for virally induced fragility but not necessarily U2 snRNA transcription per se.

9

Jiang, Chong, e Daiqing Liao. "Striking Bimodal Methylation of the Repeat Unit of the Tandem Array Encoding Human U2 snRNA (the RNU2 Locus)". Genomics 62, n.º 3 (dezembro de 1999): 508–18. http://dx.doi.org/10.1006/geno.1999.6052.

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10

Pavelitz, T., L. Rusché, A. G. Matera, J. M. Scharf e A. M. Weiner. "Concerted evolution of the tandem array encoding primate U2 snRNA occurs in situ, without changing the cytological context of the RNU2 locus." EMBO Journal 14, n.º 1 (janeiro de 1995): 169–77. http://dx.doi.org/10.1002/j.1460-2075.1995.tb06987.x.

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Bailey, Arnold D., Thomas Pavelitz e Alan M. Weiner. "The Microsatellite Sequence (CT)n · (GA)nPromotes Stable Chromosomal Integration of Large Tandem Arrays of Functional Human U2 Small Nuclear RNA Genes". Molecular and Cellular Biology 18, n.º 4 (1 de abril de 1998): 2262–71. http://dx.doi.org/10.1128/mcb.18.4.2262.

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ABSTRACT The multigene family encoding human U2 small nuclear RNA (snRNA) is organized as a single large tandem array containing 5 to 25 copies of a 6.1-kb repeat unit (the RNU2 locus). Remarkably, each of the repeat units within an individual U2 tandem array appears to be identical except for an irregular dinucleotide tract, known as the CT microsatellite, which exhibits minor length and sequence polymorphism. Using a somatic cell genetic assay, we previously noticed that the CT microsatellite appeared to stabilize artificial tandem arrays of U2 snRNA genes. We now demonstrate that the CT microsatellite is required to establish large tandem arrays of transcriptionally active U2 genes, increasing both the average and maximum size of the resulting arrays. In contrast, the CT microsatellite has no effect on the average or maximal size of artificial arrays containing transcriptionally inactive U2 genes that lack key promoter elements. Our data reinforce the connection between recombination and transcription. Active U2 transcription interferes with establishment or maintenance of the U2 tandem array, and the CT microsatellite opposes these effects, perhaps by binding GAGA or GAGA-related factors which alter local chromatin structure. We speculate that the mechanisms responsible for maintenance of tandem arrays containing active promoters may differ from those that maintain tandem arrays of transcriptionally inactive sequences.

12

Liao, D. "Concerted evolution of the tandemly repeated genes encoding human U2 snRNA (the RNU2 locus) involves rapid intrachromosomal homogenization and rare interchromosomal gene conversion". EMBO Journal 16, n.º 3 (1 de fevereiro de 1997): 588–98. http://dx.doi.org/10.1093/emboj/16.3.588.

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13

Bartsch, Detlef, Norman Gercke, Konstantin Strauch, Ronja Wieboldt, Elvira Matthäi, Vinona Wagner, Susanne Rospleszcz et al. "The Combination of MiRNA-196b, LCN2, and TIMP1 is a Potential Set of Circulating Biomarkers for Screening Individuals at Risk for Familial Pancreatic Cancer". Journal of Clinical Medicine 7, n.º 10 (20 de setembro de 2018): 295. http://dx.doi.org/10.3390/jcm7100295.

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Individuals at risk (IAR) of familial pancreatic cancer (FPC) are good candidates for screening. Unfortunately, neither reliable imaging modalities nor biomarkers are available to detect high-grade precursor lesions or early cancer. Circulating levels of candidate biomarkers LCN2, TIMP1, Glypican-1, RNU2-1f, and miRNA-196b were analyzed in 218 individuals with sporadic pancreatic ductal adenocarcinoma (PDAC, n = 50), FPC (n = 20), chronic pancreatitis (n = 10), IAR with relevant precursor lesions (n = 11) or non-relevant lesions (n = 5), 20 controls, and IAR with (n = 51) or without (n = 51) lesions on pancreatic imaging. In addition, corresponding duodenal juice samples were analyzed for Glypican-1 (n = 144) enrichment and KRAS mutations (n = 123). The panel miR-196b/LCN2/TIMP1 could distinguish high-grade lesions and stage I PDAC from controls with absolute specificity and sensitivity. In contrast, Glypican-1 enrichment in serum exosomes and duodenal juice was not diagnostic. KRAS mutations in duodenal juice were detected in 9 of 12 patients with PDAC and only 4 of 9 IAR with relevant precursor lesions. IAR with lesions on imaging had elevated miR-196b/LCN2/TIMP1 levels (p = 0.0007) and KRAS mutations in duodenal juice (p = 0.0004) significantly more often than IAR without imaging lesions. The combination miR-196b/LCN2/TIMP1 might be a promising biomarker set for the detection of high-grade PDAC precursor lesions in IAR of FPC families.

14

Pavelitz, T. "Concerted evolution of the tandem array encoding primate U2 snRNA (the RNU2 locus) is accompanied by dramatic remodeling of the junctions with flanking chromosomal sequences". EMBO Journal 18, n.º 13 (1 de julho de 1999): 3783–92. http://dx.doi.org/10.1093/emboj/18.13.3783.

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15

MacArthur, Holly L., Munna L. Agarwal e Silvia Bacchetti. "Induction of fragility at the human RNU2 locus by cytosine arabinoside is dependent upon a transcriptionally competent U2 small nuclear RNA gene and the expression of p53". Somatic Cell and Molecular Genetics 23, n.º 6 (novembro de 1997): 379–89. http://dx.doi.org/10.1007/bf02673748.

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16

Schuurman, H. J., H. P. Hougen e H. van Loveren. "The rnu (Rowett Nude) and rnuN (nznu, New Zealand Nude) Rat: An Update". ILAR Journal 34, n.º 1-2 (1 de janeiro de 1992): 3–12. http://dx.doi.org/10.1093/ilar.34.1-2.3.

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17

Li, Zengji, Arnold D. Bailey, Jacob Buchowski e Alan M. Weiner. "A Tandem Array of Minimal U1 Small Nuclear RNA Genes Is Sufficient To Generate a New Adenovirus Type 12Inducible Chromosome Fragile Site". Journal of Virology 72, n.º 5 (1 de maio de 1998): 4205–11. http://dx.doi.org/10.1128/jvi.72.5.4205-4211.1998.

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ABSTRACT Infection of human cells with adenovirus serotype 12 (Ad12) induces metaphase fragility of four, and apparently only four, chromosomal loci. Surprisingly, each of these four loci corresponds to a cluster of genes encoding a small abundant structural RNA: the RNU1and RNU2 loci contain tandemly repeated genes encoding U1 and U2 small nuclear RNAs (snRNAs), respectively; the PSU1locus is a cluster of degenerate U1 genes; and the RN5Slocus contains the tandemly repeated genes encoding 5S rRNA. These observations suggested that high local levels of transcription, in combination with Ad12 early functions, can interfere with metaphase chromatin packing. In support of this hypothesis, we and others found that an artificial tandem array of transcriptionally active, but not inactive, U2 snRNA genes would generate a novel Ad12-inducible fragile site. Although U1 and U2 snRNA are both transcribed by RNA polymerase II and share similar enhancer, promoter, and terminator signals, the human U1 promoter is clearly more complex than that of U2. In addition, the natural U1 tandem repeat unit exceeds 45 kb, whereas the U2 tandem repeat unit is only 6.1 kb. We therefore asked whether an artificial array of minimal U1 genes would also generate a novel Ad12-inducible fragile site. The exogenous U1 genes were marked by an innocuous U72C point mutation within the U1 coding region so that steady-state levels of U1 snRNA derived from the artificial array could be quantified by a simple primer extension assay. We found that the minimal U1 genes were efficiently expressed and were as effective as minimal U2 genes in generating a novel Ad12-inducible fragile site. Thus, despite significant differences in promoter architecture and overall gene organization, the active U1 transcription units suffice to generate a new virally inducible fragile site.

18

Liao, Daiqing, e Alan M. Weiner. "Concerted Evolution of the Tandemly Repeated Genes Encoding Primate U2 Small Nuclear RNA (the RNU2 Locus) Does Not Prevent Rapid Diversification of the (CT)n · (GA)n Microsatellite Embedded within the U2 Repeat Unit". Genomics 30, n.º 3 (dezembro de 1995): 583–93. http://dx.doi.org/10.1006/geno.1995.1280.

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19

Tatsuta, Masahiro, Hiroyuki Mizumoto, Masanori Kaido, Kazuyuki Mise e Tetsuro Okuno. "The Red Clover Necrotic Mosaic Virus RNA2 trans-Activator Is Also a cis-Acting RNA2 Replication Element". Journal of Virology 79, n.º 2 (15 de janeiro de 2005): 978–86. http://dx.doi.org/10.1128/jvi.79.2.978-986.2005.

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ABSTRACT The expression of the coat protein gene requires RNA-mediated trans-activation of subgenomic RNA synthesis in Red clover necrotic mosaic virus (RCNMV), the genome of which consists of two positive-strand RNAs, RNA1 and RNA2. The trans-acting RNA element required for subgenomic RNA synthesis from RNA1 has been mapped previously to the protein-coding region of RNA2, whereas RNA2 is not required for the replication of RNA1. In this study, we investigated the roles of the protein-coding region in RNA2 replication by analyzing the replication competence of RNA2 mutants containing deletions or nucleotide substitutions. Our results indicate that the same stem-loop structure (SL2) that functions as a trans-activator for RNA-mediated coat protein expression is critically required for the replication of RNA2 itself. Interestingly, however, disruption of the RNA-RNA interaction by nucleotide substitutions in the region of RNA1 corresponding to the SL2 loop of RNA2 does not affect RNA2 replication, indicating that the RNA-RNA interaction is not required for RNA2 replication. Further mutational analysis showed that, in addition to the stem-loop structure itself, nucleotide sequences in the stem and in the loop of SL2 are important for the replication of RNA2. These findings suggest that the structure and nucleotide sequence of SL2 in RNA2 play multiple roles in the virus life cycle.

20

Yang, Jie, Fan Yu, Jinlei Guan, Tao Wang, Changjiang Liu, Yuxiang Wang, Guangjie Liu e Shuchai Zhu. "Knockdown of RNF2 enhances the radiosensitivity of squamous cell carcinoma in lung". Biochemistry and Cell Biology 97, n.º 5 (outubro de 2019): 589–99. http://dx.doi.org/10.1139/bcb-2018-0252.

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A previous study has reported that knockdown of RING finger protein 2 (RNF2) increases the radiosensitivity of esophageal cancer cells both in vitro and in vivo. However, the effect of RNF2 knockdown on radiosensitivity in squamous cell carcinoma (SqCC) remains unknown. For this, NCI-H226 and SK-MES-1 cells were exposed to X-ray irradiation and then RNF2 levels were determined. RNF2 was knocked-down and stable transfectants were selected. Radiosensitivity, cell proliferation, apoptosis, cell cycle, and γ-H2AX foci formation were evaluated. Interaction among ataxia telangiectasia mutated protein (ATM), mediator of DNA damage checkpoint 1 (MDC1), and H2AX were examined. Xenograft models were used to explore the effect of RNF2 knockdown on radiosensitivity in vivo. The results showed that RNF2 expression was significantly increased by X-ray irradiation. RNF2 knockdown combined with X-ray irradiation markedly inhibited cell proliferation, caused cell cycle arrest at the G1 phase, and induced cell apoptosis. In addition, RNF2 knockdown enhanced the radiosensitivity of SqCC cells, inhibited irradiation-induced γ-H2AX foci formation, and impaired the interactions among ATM, MDC1, and H2AX. Furthermore, combination of RNF2 knockdown and X-ray irradiation suppressed tumor growth and promoted tumor cell apoptosis in vivo. RNF2 may be a new therapeutic target to enhance the radiosensitivity of SqCC cells in lung.

21

Mizumoto, Hiroyuki, Hiro-oki Iwakawa, Masanori Kaido, Kazuyuki Mise e Tetsuro Okuno. "Cap-Independent Translation Mechanism of Red Clover Necrotic Mosaic Virus RNA2 Differs from That of RNA1 and Is Linked to RNA Replication". Journal of Virology 80, n.º 8 (15 de abril de 2006): 3781–91. http://dx.doi.org/10.1128/jvi.80.8.3781-3791.2006.

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ABSTRACT The genome of Red clover necrotic mosaic virus (RCNMV) in the genus Dianthovirus is divided into two RNA molecules of RNA1 and RNA2, which have no cap structure at the 5′ end and no poly(A) tail at the 3′ end. The 3′ untranslated region (3′ UTR) of RCNMV RNA1 contains an essential RNA element (3′TE-DR1), which is required for cap-independent translation. In this study, we investigated a cap-independent translational mechanism of RNA2 using a firefly luciferase (Luc) gene expression assay system in cowpea protoplasts and a cell-free lysate (BYL) prepared from evacuolated tobacco BY2 protoplasts. We were unable to detect cis-acting RNA sequences in RNA2 that can replace the function of a cap structure, such as the 3′TE-DR1 of RNA1. However, the uncapped reporter RNA2, RNA2-Luc, in which the Luc open reading frame (ORF) was inserted between the 5′ UTR and the movement protein ORF, was effectively translated in the presence of p27 and p88 in protoplasts in which RNA2-Luc was replicated. Time course experiments in protoplasts showed that the translational activity of RNA2-Luc did not reflect the amount of RNA2. Mutations in cis-acting RNA replication elements of RNA2 abolished the cap-independent translational activity of RNA2-Luc, suggesting that the translational activity of RNA2-Luc is coupled to RNA replication. Our results show that the translational mechanism differs between two segmented genomic RNAs of RCNMV. We present a model in which only RNA2 that is generated de novo through the viral RNA replication machinery functions as mRNA for translation.

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Elledge, S. J., e R. W. Davis. "DNA damage induction of ribonucleotide reductase." Molecular and Cellular Biology 9, n.º 11 (novembro de 1989): 4932–40. http://dx.doi.org/10.1128/mcb.9.11.4932.

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RNR2 encodes the small subunit of ribonucleotide reductase, the enzyme that catalyzes the first step in the pathway for the production of deoxyribonucleotides needed for DNA synthesis. RNR2 is a member of a group of genes whose activities are cell cycle regulated and that are transcriptionally induced in response to the stress of DNA damage. An RNR2-lacZ fusion was used to further characterize the regulation of RNR2 and the pathway responsible for its response to DNA damage. beta-Galactosidase activity in yeast strains containing the RNR2-lacZ fusion was inducible in response to DNA-damaging agents (UV light, 4-nitroquinoline-1-oxide [4-NQO], and methyl methanesulfonate [MMS]) and agents that block DNA replication (hydroxyurea [HU] and methotrexate) but not heat shock. When MATa cells were arrested in G1 by alpha-factor, RNR2 mRNA was still inducible by DNA damage, indicating that the observed induction can occur outside of S phase. In addition, RNR2 induction was not blocked by the presence of cycloheximide and is therefore likely to be independent of protein synthesis. A mutation, rnr2-314, was found to confer hypersensitivity to HU and increased sensitivity to MMS. In rnr2-314 mutant strains, the DNA damage stress response was found to be partially constitutive as well as hypersensitive to induction by HU but not MMS. The induction properties of RNR2 were examined in a rad4-2 mutant background; in this genetic background, RNR2 was hypersensitive to induction by 4-NQO but not MMS. Induction of the RNR2-lacZ fusion in a RAD(+) strain in response to 4-NQO was not enhanced by the presence of an equal number of rad4-2 cells that lacked the fusion, implying that the DNA damage stress response in cell autonomous.

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Chen, Jianbo, Amine Noueiry e Paul Ahlquist. "Brome Mosaic Virus Protein 1a Recruits Viral RNA2 to RNA Replication through a 5′ Proximal RNA2 Signal". Journal of Virology 75, n.º 7 (1 de abril de 2001): 3207–19. http://dx.doi.org/10.1128/jvi.75.7.3207-3219.2001.

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ABSTRACT Brome mosaic virus (BMV), a positive-strand RNA virus in the alphavirus-like superfamily, encodes two RNA replication factors. Membrane-associated 1a protein contains a helicase-like domain and RNA capping functions. 2a, which is targeted to membranes by 1a, contains a central polymerase-like domain. In the absence of 2a and RNA replication, 1a acts through an intergenic replication signal in BMV genomic RNA3 to stabilize RNA3 and induce RNA3 to associate with cellular membrane. Multiple results imply that 1a-induced RNA3 stabilization reflects interactions involved in recruiting RNA3 templates into replication. To determine if 1a had similar effects on another BMV RNA replication template, we constructed a plasmid expressing BMV genomic RNA2 in vivo. In vivo-expressed RNA2 templates were replicated upon expression of 1a and 2a. In the absence of 2a, 1a stabilized RNA2 and induced RNA2 to associate with membrane. Deletion analysis demonstrated that 1a-induced membrane association of RNA2 was mediated by sequences in the 5′-proximal third of RNA2. The RNA2 5′ untranslated region was sufficient to confer 1a-induced membrane association on a nonviral RNA. However, sequences in the N-terminal region of the 2a open reading frame enhanced 1a responsiveness of RNA2 and a chimeric RNA. A 5′-terminal RNA2 stem-loop important for RNA2 replication was essential for 1a-induced membrane association of RNA2 and, like the 1a-responsive RNA3 intergenic region, contained a required box B motif corresponding to the TΨC stem-loop of host tRNAs. The level of 1a-induced membrane association of various RNA2 mutants correlated well with their abilities to serve as replication templates. These results support and expand the conclusion that 1a-induced BMV RNA stabilization and membrane association reflect early, 1a-mediated steps in viral RNA replication.

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Elledge, S. J., e R. W. Davis. "DNA damage induction of ribonucleotide reductase". Molecular and Cellular Biology 9, n.º 11 (novembro de 1989): 4932–40. http://dx.doi.org/10.1128/mcb.9.11.4932-4940.1989.

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RNR2 encodes the small subunit of ribonucleotide reductase, the enzyme that catalyzes the first step in the pathway for the production of deoxyribonucleotides needed for DNA synthesis. RNR2 is a member of a group of genes whose activities are cell cycle regulated and that are transcriptionally induced in response to the stress of DNA damage. An RNR2-lacZ fusion was used to further characterize the regulation of RNR2 and the pathway responsible for its response to DNA damage. beta-Galactosidase activity in yeast strains containing the RNR2-lacZ fusion was inducible in response to DNA-damaging agents (UV light, 4-nitroquinoline-1-oxide [4-NQO], and methyl methanesulfonate [MMS]) and agents that block DNA replication (hydroxyurea [HU] and methotrexate) but not heat shock. When MATa cells were arrested in G1 by alpha-factor, RNR2 mRNA was still inducible by DNA damage, indicating that the observed induction can occur outside of S phase. In addition, RNR2 induction was not blocked by the presence of cycloheximide and is therefore likely to be independent of protein synthesis. A mutation, rnr2-314, was found to confer hypersensitivity to HU and increased sensitivity to MMS. In rnr2-314 mutant strains, the DNA damage stress response was found to be partially constitutive as well as hypersensitive to induction by HU but not MMS. The induction properties of RNR2 were examined in a rad4-2 mutant background; in this genetic background, RNR2 was hypersensitive to induction by 4-NQO but not MMS. Induction of the RNR2-lacZ fusion in a RAD(+) strain in response to 4-NQO was not enhanced by the presence of an equal number of rad4-2 cells that lacked the fusion, implying that the DNA damage stress response in cell autonomous.

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Kazibwe, Zakayo, Junmarie Soto-Burgos, Gustavo C. MacIntosh e Diane C. Bassham. "TOR mediates the autophagy response to altered nucleotide homeostasis in an RNase mutant". Journal of Experimental Botany 71, n.º 22 (9 de setembro de 2020): 6907–20. http://dx.doi.org/10.1093/jxb/eraa410.

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Abstract The Arabidopsis thaliana T2 family endoribonuclease RNS2 localizes to the vacuole and functions in rRNA degradation. Loss of RNS2 activity impairs rRNA turnover and leads to constitutive autophagy, a process for degradation of cellular components. Autophagy is normally activated during environmental stress and is important for stress tolerance and homeostasis. Here we show that restoration of cytosolic purine nucleotide levels rescues the constitutive autophagy phenotype of rns2-2 seedlings, whereas inhibition of purine synthesis induces autophagy in wild-type seedlings. rns2-2 seedlings have reduced activity of the target of rapamycin (TOR) kinase complex, a negative regulator of autophagy, and this phenotype is rescued by addition of inosine to increase purine levels. Activation of TOR in rns2-2 by exogenous auxin blocks the enhanced autophagy, indicating a possible involvement of the TOR signaling pathway in the activation of autophagy in the rns2-2 mutant. Our data suggest a model in which loss of rRNA degradation in rns2-2 leads to a reduction in cytoplasmic nucleotide concentrations, which in turn inhibits TOR activity, leading to activation of autophagy to restore homeostasis.

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TANAKA, Hiroko, Hirotada FUJITA, Hironori KATOH, Kazutoshi MORI e Manabu NEGISHI. "Vps4-A (vacuolar protein sorting 4-A) is a binding partner for a novel Rho family GTPase, Rnd2". Biochemical Journal 365, n.º 2 (15 de julho de 2002): 349–53. http://dx.doi.org/10.1042/bj20020062.

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Rho family GTPases are implicated in a variety of biological activities, including endocytic vesicle trafficking. Rnd2 is a new member of Rho family GTPases, but its biological functions are not known. In the present study, we have performed a yeast two-hybrid screening using Rnd2 as bait and revealed that Rnd2 binds specifically to Vps4-A (where Vsp4-A is vacuolar protein sorting 4-A), a member of the AAA ATPase family and a central regulator for early endosome trafficking. This interaction was determined by the yeast two-hybrid system, in vitro binding and co-immunoprecipitation studies. Vps4-A associated with both guanosine 5′-[β-thio]triphosphate-bound active and guanosine 5′-[β-thio]diphosphate-bound inactive forms of Rnd2. An ATPase-defective Vps4-A mutant, Vps4-AE228Q, expressed in HeLa cells was accumulated in the early endosomes. When Rnd2 was co-expressed with Vps4-AE228Q, Rnd2 was recruited to the Vps4-A-bound early endosomes. These results suggest that Rnd2 is involved in the regulation of endosomal trafficking via direct binding to Vps4-A.

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Lee, Sun-Joo, Dongwon Choi, Hyangshuk Rhim e Seongman Kang. "E3 ubiquitin ligase RNF2 interacts with the S6′ proteasomal ATPase subunit and increases the ATP hydrolysis activity of S6′". Biochemical Journal 389, n.º 2 (5 de julho de 2005): 457–63. http://dx.doi.org/10.1042/bj20041982.

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We reported previously that the human RNF2 (RING finger protein 2) protein is an E3 ubiquitin ligase that interacts with the human ubiquitin-conjugating enzyme Hip-2/hE2-25K. In the present study, we show that RNF2 interacts with S6′ ATPase, a subunit of the proteasomal 19 S regulatory complex. S6′ interacts with RNF2 through its N-terminal RING domain, and RNF2 interacts with S6′ through its C-terminal region. Interestingly, the RNF2-S6′ interaction increases the ATP hydrolysis activity of the S6′ protein. Moreover, S6′ ATPase activity is highly increased in the presence of ubiquitinated proteins. The present study suggests that the E3 ubiquitin ligase RNF2 might have a dual function: facilitating the ubiquitination of its target substrates and recruiting the substrates to the proteasome. Furthermore, ATP hydrolysis in the E3/proteasome complex might act as an important signal for the protein degradation pathway.

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Elledge, S. J., e R. W. Davis. "Identification of the DNA damage-responsive element of RNR2 and evidence that four distinct cellular factors bind it." Molecular and Cellular Biology 9, n.º 12 (dezembro de 1989): 5373–86. http://dx.doi.org/10.1128/mcb.9.12.5373.

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The RNR2 gene encodes the small subunit of ribonucleotide reductase, the enzyme that catalyzes the first step in the pathway for the production of the deoxyribonucleotides needed for DNA synthesis. Transcription of this gene is induced approximately 20-fold in response to environmental stimuli that damage DNA or block DNA replication. Deletion and subcloning analysis identified two, and possibly three, upstream activating sequences (UAS) and one repressing (URS) element in the RNR2 regulatory region. A 42-base-pair (bp) fragment from this region was found to be necessary for proper regulation of RNR2 and to be capable of conferring DNA damage inducibility upon a heterologous promoter. This fragment contained both positively and negatively acting sequences. Four DNA-binding factors interacted with the RNR2 regulatory region. One factor was identified as the GRF1 protein, the product of the RAP1 gene. GRF1 bound to the UAS2 element of RNR2, which was found to be directly adjacent to the 42-bp fragment. UAS2 activity was repressed by the 42-bp fragment. Three other factors bound to the 42-bp fragment; one of these factors, RRF3, had a second binding site in the RNR2 promoter. These factors are likely to mediate the response of RNR2 to DNA damage.

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Olsthoorn, R. C. L., A. Bruyere, A. Dzianott e J. J. Bujarski. "RNA Recombination in Brome Mosaic Virus: Effects of Strand-Specific Stem-Loop Inserts". Journal of Virology 76, n.º 24 (15 de dezembro de 2002): 12654–62. http://dx.doi.org/10.1128/jvi.76.24.12654-12662.2002.

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ABSTRACT A model system of a single-stranded trisegment Brome mosaic bromovirus (BMV) was used to analyze the mechanism of homologous RNA recombination. Elements capable of forming strand-specific stem-loop structures were inserted at the modified 3′ noncoding regions of BMV RNA3 and RNA2 in either positive or negative orientations, and various combinations of parental RNAs were tested for patterns of the accumulating recombinant RNA3 components. The structured negative-strand stem-loops that were inserted in both RNA3 and RNA2 reduced the accumulation of RNA3-RNA2 recombinants to a much higher extent than those in positive strands or the unstructured stem-loop inserts in either positive or negative strands. The use of only one parental RNA carrying the stem-loop insert reduced the accumulation of RNA3-RNA2 recombinants even further, but only when the stem-loops were in negative strands of RNA2. We assume that the presence of a stable stem-loop downstream of the landing site on the acceptor strand (negative RNA2) hampers the reattachment and reinitiation processes. Besides RNA3-RNA2 recombinants, the accumulation of nontargeted RNA3-RNA1 and RNA3-RNA3 recombinants were observed. Our results provide experimental evidence that homologous recombination between BMV RNAs more likely occurs during positive- rather than negative-strand synthesis.

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Elledge, S. J., e R. W. Davis. "Identification of the DNA damage-responsive element of RNR2 and evidence that four distinct cellular factors bind it". Molecular and Cellular Biology 9, n.º 12 (dezembro de 1989): 5373–86. http://dx.doi.org/10.1128/mcb.9.12.5373-5386.1989.

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The RNR2 gene encodes the small subunit of ribonucleotide reductase, the enzyme that catalyzes the first step in the pathway for the production of the deoxyribonucleotides needed for DNA synthesis. Transcription of this gene is induced approximately 20-fold in response to environmental stimuli that damage DNA or block DNA replication. Deletion and subcloning analysis identified two, and possibly three, upstream activating sequences (UAS) and one repressing (URS) element in the RNR2 regulatory region. A 42-base-pair (bp) fragment from this region was found to be necessary for proper regulation of RNR2 and to be capable of conferring DNA damage inducibility upon a heterologous promoter. This fragment contained both positively and negatively acting sequences. Four DNA-binding factors interacted with the RNR2 regulatory region. One factor was identified as the GRF1 protein, the product of the RAP1 gene. GRF1 bound to the UAS2 element of RNR2, which was found to be directly adjacent to the 42-bp fragment. UAS2 activity was repressed by the 42-bp fragment. Three other factors bound to the 42-bp fragment; one of these factors, RRF3, had a second binding site in the RNR2 promoter. These factors are likely to mediate the response of RNR2 to DNA damage.

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Last, R. L., e J. L. Woolford. "Identification and nuclear localization of yeast pre-messenger RNA processing components: RNA2 and RNA3 proteins." Journal of Cell Biology 103, n.º 6 (1 de dezembro de 1986): 2103–12. http://dx.doi.org/10.1083/jcb.103.6.2103.

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Temperature-sensitive mutations in the RNA2 through RNA11 genes of yeast prevent the processing of nuclear pre-mRNAs. We have raised antisera that detect the RNA2 and RNA3 proteins in immunoblots of extracts of yeast containing high copy number RNA2 and RNA3 plasmids. Subcellular fractionation of yeast cells that overproduce the RNA2 and RNA3 proteins has revealed that these proteins are enriched in nuclear fractions. Indirect immunofluorescence results have indicated that these proteins are localized in yeast nuclei. These localization results are consistent with the fact that these genes have a role in processing yeast pre-mRNA.

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Black, Joshua C., e Johnathan R. Whetstine. "RNF2 E3 or Not to E3: Dual Roles of RNF2 Overexpression in Melanoma". Cancer Discovery 5, n.º 12 (dezembro de 2015): 1241–43. http://dx.doi.org/10.1158/2159-8290.cd-15-1285.

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Hu, C. C., e S. A. Ghabrial. "Molecular Evidence That Strain BV-15 of Peanut Stunt Cucumovirus Is a Reassortant Between Subgroup I and II Strains". Phytopathology® 88, n.º 2 (fevereiro de 1998): 92–97. http://dx.doi.org/10.1094/phyto.1998.88.2.92.

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In Northern hybridization assays, RNA1 of peanut stunt virus (PSV) strain BV-15 hybridized strongly with a cloned cDNA probe to RNA1 from strain PSV-W (subgroup II). Cloned probes to PSV-W RNA2 and RNA3, however, did not hybridize with the corresponding RNAs from strain BV-15. The complete nucleotide sequence of PSV-BV-15 RNA2 has been determined, and sequence comparison analysis showed that it is closely related to PSV subgroup I strains; the percent nucleotide sequence identity between PSV-BV-15 RNA2 and RNA2 sequences from PSV subgroup I and II strains were 90.5 and 75%, respectively. The possibility that PSV-BV-15 RNA2 may contain short regions derived from a subgroup II strain (i.e., represent a mosaic structure indicative of recombination) was investigated. Results indicated, however, that the entire PSV-BV-15 RNA2 sequence is derived from a subgroup I strain. PSV-BV-15 RNA3 has previously been shown to belong to subgroup I strains. These results thus establish that PSV strain BV-15 is a natural reassortant between PSV subgroups I and II strains. A reverse transcription-po-lymerase chain reaction assay is proposed for differentiating between this reassortant strain and PSV strains in subgroups I and II.

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Vega, Aracelly, e Heriberto Franco. "Productividad y calidad de los cuerpos fructíferos de los hongos comestibles Pleurotus pulmonarius RN2 y P. djamor RN81 y RN82 cultivados sobre sustratos lignocelulósicos". Información tecnológica 24, n.º 1 (2013): 69–78. http://dx.doi.org/10.4067/s0718-07642013000100009.

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Morel, Etienne, Nicolas Dupont e Patrice Codogno. "Autophagy regulation: RNF2 targets AMBRA1". Cell Research 24, n.º 9 (8 de agosto de 2014): 1029–30. http://dx.doi.org/10.1038/cr.2014.105.

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Paudel, Dinesh Babu, e Hélène Sanfaçon. "Mapping of sequences in the 5’ region and 3’ UTR of tomato ringspot virus RNA2 that facilitate cap-independent translation of reporter transcripts in vitro". PLOS ONE 16, n.º 4 (9 de abril de 2021): e0249928. http://dx.doi.org/10.1371/journal.pone.0249928.

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Tomato ringspot virus (ToRSV, genus Nepovirus, family Secoviridae, order Picornavirales) is a bipartite positive-strand RNA virus, with each RNA encoding one large polyprotein. ToRSV RNAs are linked to a 5’-viral genome-linked protein (VPg) and have a 3’ polyA tail, suggesting a non-canonical cap-independent translation initiation mechanism. The 3’ untranslated regions (UTRs) of RNA1 and RNA2 are unusually long (~1.5 kb) and share several large stretches of sequence identities. Several putative in-frame start codons are present in the 5’ regions of the viral RNAs, which are also highly conserved between the two RNAs. Using reporter transcripts containing the 5’ region and 3’ UTR of the RNA2 of ToRSV Rasp1 isolate (ToRSV-Rasp1) and in vitro wheat germ extract translation assays, we provide evidence that translation initiates exclusively at the first AUG, in spite of a poor codon context. We also show that both the 5’ region and 3’ UTR of RNA2 are required for efficient cap-independent translation of these transcripts. We identify translation-enhancing elements in the 5’ proximal coding region of the RNA2 polyprotein and in the RNA2 3’ UTR. Cap-dependent translation of control reporter transcripts was inhibited when RNAs consisting of the RNA2 3’ UTR were supplied in trans. Taken together, our results suggest the presence of a CITE in the ToRSV-Rasp1 RNA2 3’ UTR that recruits one or several translation factors and facilitates efficient cap-independent translation together with the 5’ region of the RNA. Non-overlapping deletion mutagenesis delineated the putative CITE to a 200 nts segment (nts 773–972) of the 1547 nt long 3’ UTR. We conclude that the general mechanism of ToRSV RNA2 translation initiation is similar to that previously reported for the RNAs of blackcurrant reversion virus, another nepovirus. However, the position, sequence and predicted structures of the translation-enhancing elements differed between the two viruses.

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Lin, Junyan, Ahmed Khamis Ali, Ping Chen, Said Ghabrial, John Finer, Anne Dorrance, Peg Redinbaugh e Feng Qu. "A stem–loop structure in the 5′ untranslated region of bean pod mottle virus RNA2 is specifically required for RNA2 accumulation". Journal of General Virology 94, n.º 6 (1 de junho de 2013): 1415–20. http://dx.doi.org/10.1099/vir.0.051755-0.

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Bean pod mottle virus (BPMV) is a bipartite, positive-sense (+) RNA plant virus of the family Secoviridae. Its RNA1 encodes all proteins needed for genome replication and is capable of autonomous replication. By contrast, BPMV RNA2 must utilize RNA1-encoded proteins for replication. Here, we sought to identify RNA elements in RNA2 required for its replication. The exchange of 5′ untranslated regions (UTRs) between genome segments revealed an RNA2-specific element in its 5′ UTR. Further mapping localized a 66 nucleotide region that was predicted to fold into an RNA stem–loop structure, designated SLC. Additional functional analysis indicated the importance of the middle portion of the stem and an adjacent two-base mismatch. This is the first report of a cis-acting RNA element in RNA2 of a bipartite secovirus.

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Nagy, Peter D., e Jozef J. Bujarski. "Silencing Homologous RNA Recombination Hot Spots with GC-Rich Sequences in Brome Mosaic Virus". Journal of Virology 72, n.º 2 (1 de fevereiro de 1998): 1122–30. http://dx.doi.org/10.1128/jvi.72.2.1122-1130.1998.

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ABSTRACT It has been observed that AU-rich sequences form homologous recombination hot spots in brome mosaic virus (BMV), a tripartite positive-stranded RNA virus of plants (P. D. Nagy and J. J. Bujarski, J. Virol. 71:3799–3810, 1997). To study the effect of GC-rich sequences on the recombination hot spots, we inserted 30-nucleotide-long GC-rich sequences downstream of AU-rich homologous recombination hot spot regions in parental BMV RNAs (RNA2 and RNA3). Although these insertions doubled the length of sequence identity in RNA2 and RNA3, the incidence of homologous RNA2 and RNA3 recombination was reduced markedly. Four different, both highly structured and nonstructured downstream GC-rich sequences had a similar “homologous recombination silencing” effect on the nearby hot spots. The GC-rich sequence-mediated recombination silencing mapped to RNA2, as it was observed when the GC-rich sequence was inserted at downstream locations in both RNA2 and RNA3 or only in the RNA2 component. On the contrary, when the downstream GC-rich sequence was present only in the RNA3 component, it increased the incidence of homologous recombination. In addition, upstream insertions of similar GC-rich sequences increased the incidence of homologous recombination within downstream hot spot regions. Overall, this study reveals the complex nature of homologous recombination in BMV, where sequences flanking the common hot spot regions affect recombination frequency. A replicase-driven template-switching model is presented to explain recombination silencing by GC-rich sequences.

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Luo, Xi, Kelly Schoch, Sharayu V. Jangam, Venkata Hemanjani Bhavana, Hillary K. Graves, Sujay Kansagra, Joan M. Jasien et al. "Rare deleterious de novo missense variants in Rnf2/Ring2 are associated with a neurodevelopmental disorder with unique clinical features". Human Molecular Genetics 30, n.º 14 (16 de abril de 2021): 1283–92. http://dx.doi.org/10.1093/hmg/ddab110.

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Abstract The Polycomb group (PcG) gene RNF2 (RING2) encodes a catalytic subunit of the Polycomb repressive complex 1 (PRC1), an evolutionarily conserved machinery that post-translationally modifies chromatin to maintain epigenetic transcriptional repressive states of target genes including Hox genes. Here, we describe two individuals, each with rare de novo missense variants in RNF2. Their phenotypes include intrauterine growth retardation, severe intellectual disabilities, behavioral problems, seizures, feeding difficulties and dysmorphic features. Population genomics data suggest that RNF2 is highly constrained for loss-of-function (LoF) and missense variants, and both p.R70H and p.S82R variants have not been reported to date. Structural analyses of the two alleles indicate that these changes likely impact the interaction between RNF2 and BMI1, another PRC1 subunit or its substrate Histone H2A, respectively. Finally, we provide functional data in Drosophila that these two missense variants behave as LoF alleles in vivo. The evidence provide support for deleterious alleles in RNF2 being associated with a new and recognizable genetic disorder. This tentative gene-disease association in addition to the 12 previously identified disorders caused by PcG genes attests to the importance of these chromatin regulators in Mendelian disorders.

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Last, Robert L., Janine R. Maddock e John L. Woolford. "Evidence for Related Functions of the RNA Genes of Saccharomyces cerevisiae". Genetics 117, n.º 4 (1 de dezembro de 1987): 619–31. http://dx.doi.org/10.1093/genetics/117.4.619.

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ABSTRACT The yeast genes RNA2-RNA11 are necessary for splicing of nuclear intron-containing pre-mRNAs. We investigated the relationships among these genes by asking whether increased expression of one RNA gene leads to suppression of the temperature-sensitive lethality of a mutation in any other RNA gene. The presence of extra plasmid-borne copies of the RNA3 gene relieves the lethality of temperature-sensitive rna4 mutations. A region of the yeast genome (SRN2) is described that suppresses temperature-sensitive rna2 mutations when it is present on either medium or high-copy number plasmids. Neither suppression occurs via a bypass of RNA gene function since null alleles of rna2 and rna4 are not suppressed by elevated dosage of SRN2 and RNA3, respectively. These results suggest that the SRN2 and RNA2 gene products have related functions, as do the RNA3 and RNA4 gene products.

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Hurd, H. K., e J. W. Roberts. "Upstream regulatory sequences of the yeast RNR2 gene include a repression sequence and an activation site that binds the RAP1 protein." Molecular and Cellular Biology 9, n.º 12 (dezembro de 1989): 5359–72. http://dx.doi.org/10.1128/mcb.9.12.5359.

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The small subunit of ribonucleotide reductase in Saccharomyces cerevisiae (RNR2) was induced 3- to 20-fold by a variety of DNA-damaging agents. Induction of the RNR2 transcript by at least one of these agents, methyl methanesulfonate, did not require protein synthesis. To identify sequences involved in the regulation of RNR2, we introduced deletions upstream of the transcription start site. Sequences required for induction were contained within a 200-base-pair region that could confer methyl methanesulfonate inducibility on the heterologous CYC1 promoter. This region contained a repression sequence and at least two positive activation sites. One of these activation sites bound RAP1, a protein known to associate with mating-type silencers and the upstream activation sequences of a number of genes. The behavior of deletions of the repression sequence suggests that induction of RNR2 may occur, at least in part, through relief of repression.

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Hurd, H. K., e J. W. Roberts. "Upstream regulatory sequences of the yeast RNR2 gene include a repression sequence and an activation site that binds the RAP1 protein". Molecular and Cellular Biology 9, n.º 12 (dezembro de 1989): 5359–72. http://dx.doi.org/10.1128/mcb.9.12.5359-5372.1989.

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The small subunit of ribonucleotide reductase in Saccharomyces cerevisiae (RNR2) was induced 3- to 20-fold by a variety of DNA-damaging agents. Induction of the RNR2 transcript by at least one of these agents, methyl methanesulfonate, did not require protein synthesis. To identify sequences involved in the regulation of RNR2, we introduced deletions upstream of the transcription start site. Sequences required for induction were contained within a 200-base-pair region that could confer methyl methanesulfonate inducibility on the heterologous CYC1 promoter. This region contained a repression sequence and at least two positive activation sites. One of these activation sites bound RAP1, a protein known to associate with mating-type silencers and the upstream activation sequences of a number of genes. The behavior of deletions of the repression sequence suggests that induction of RNR2 may occur, at least in part, through relief of repression.

43

Chen, Ying, Xiao-Yun Cao, Ying-Ni Li, Yu-Yan Qiu, Ying-Na Li, Wen Li e Hui Wang. "Reversal of cisplatin resistance by microRNA-139-5p-independent RNF2 downregulation and MAPK inhibition in ovarian cancer". American Journal of Physiology-Cell Physiology 315, n.º 2 (1 de agosto de 2018): C225—C235. http://dx.doi.org/10.1152/ajpcell.00283.2017.

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Some microRNAs (miRs) are dysregulated in cancers, and aberrant miR expression has been reported to correlate with chemoresistance of cancer cells. Therefore, the present study aims at investigating the effects of microRNA-139-5p (miR-139-5p) on cisplatin resistance of ovarian cancer (OC) with involvement of ring finger protein 2 (RNF2) and the mitogen-activated protein kinase (MAPK) signaling pathway. OC tissues were obtained from 66 primary OC patients. The cisplatin-sensitive A2780 and cisplatin-resistant A2780/DDP cell lines were collected for construction of RNF2 silencing and overexpressed plasmids. Cell vitality and apoptosis were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and annexin V-FITC/propidium iodide double-staining, respectively. Next, expression of RNF2, extracellular signal-related kinase, and p38 was determined by quantitative reverse transcription-quantitative polymerase chain reaction and Western blot analysis. Finally, the volume of xenograft tumors in BALB/c nude mice was detected. RNF2 and miR-139-5p were identified to be involved in OC. In addition, MAPK activation and RNF2 were related to cisplatin resistance of OC. miR-139-5p was downregulated in cisplatin-resistant OC tissues, and miR-139-5p overexpression could inhibit cell vitality, reduce cisplatin resistance, and promote apoptosis of OC cells. Furthermore, miR-139-5p combined with MAPK inhibitors more obviously reduced cisplatin resistance of OC. Taken together, this study demonstrated that miR-139-5p overexpression combined with inactivation of the MAPK signaling pathway can reverse the cisplatin resistance of OC by suppressing RNF2. Thus, miR-139-5p overexpression might be a future therapeutic strategy for OC.

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Venter, P. Arno, Neel K. Krishna e Anette Schneemann. "Capsid Protein Synthesis from Replicating RNA Directs Specific Packaging of the Genome of a Multipartite, Positive-Strand RNA Virus". Journal of Virology 79, n.º 10 (15 de maio de 2005): 6239–48. http://dx.doi.org/10.1128/jvi.79.10.6239-6248.2005.

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ABSTRACT Flock house virus (FHV) is a bipartite, positive-strand RNA insect virus that encapsidates its two genomic RNAs in a single virion. It provides a convenient model system for studying the principles underlying the copackaging of multipartite viral RNA genomes. In this study, we used a baculovirus expression system to determine if the uncoupling of viral protein synthesis from RNA replication affected the packaging of FHV RNAs. We found that neither RNA1 (which encodes the viral replicase) nor RNA2 (which encodes the capsid protein) were packaged efficiently when capsid protein was supplied in trans from nonreplicating RNA. However, capsid protein synthesized in cis from replicating RNA2 packaged RNA2 efficiently in the presence and absence of RNA1. These results demonstrated that capsid protein translation from replicating RNA2 is required for specific packaging of the FHV genome. This type of coupling between genome replication and translation and RNA packaging has not been observed previously. We hypothesize that RNA2 replication and translation must be spatially coordinated in FHV-infected cells to facilitate retrieval of the viral RNAs for encapsidation by newly synthesized capsid protein. Spatial coordination of RNA and capsid protein synthesis may be key to specific genome packaging and assembly in other RNA viruses.

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Miyamoto, Yuki, Tomohiro Torii, Miho Terao, Shuji Takada, Akito Tanoue, Hironori Katoh e Junji Yamauchi. "Rnd2 differentially regulates oligodendrocyte myelination at different developmental periods". Molecular Biology of the Cell 32, n.º 8 (15 de abril de 2021): 769–87. http://dx.doi.org/10.1091/mbc.e20-05-0332.

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The unique biphasic role of Rnd2, an atypical branch of the Rho family GTPases, in oligodendrocyte myelination is described. Rnd2 positively regulates myelination in the early myelinating period and negatively regulates it in the subsequent period, by changes of the Rho-kinase activities, a common downstream molecule of Rho GTPases.

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Gardner, Amy L., James K. Roche, Richard L. Guerrant e Cynthia S. Weikel. "Intestinal Cryptosporidiosis: Pathophysiologic Alterations and Specific Cellular and Humoral Immune Responses in Rnu/+ and Rnu/Rnu (Athymic) Rats". American Journal of Tropical Medicine and Hygiene 44, n.º 1 (1 de janeiro de 1991): 49–62. http://dx.doi.org/10.4269/ajtmh.1991.44.49.

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Delsante, S., R. Stifanese e G. Borzone. "Thermodynamic stability of RNi2 Laves phases". Journal of Chemical Thermodynamics 65 (outubro de 2013): 73–77. http://dx.doi.org/10.1016/j.jct.2013.05.044.

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Gao, Shuangyu, Jinda Lu, Xiaodong Cheng, Zhouhang Gu, Qiansheng Liao e Zhiyou Du. "Heterologous Replicase from Cucumoviruses can Replicate Viral RNAs, but is Defective in Transcribing Subgenomic RNA4A or Facilitating Viral Movement". Viruses 10, n.º 11 (28 de outubro de 2018): 590. http://dx.doi.org/10.3390/v10110590.

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Interspecific exchange of RNA1 or RNA2 between the cucumoviruses cucumber mosaic virus (CMV) and tomato aspermy virus (TAV) was reported to be non-viable in plants previously. Here we investigated viability of the reassortants between CMV and TAV in Nicotiana benthamiana plants by Agrobacterium-mediated viral inoculation. The reassortants were composed of CMV RNA1 and TAV RNA2 plus RNA3 replicated in the inoculated leaves, while they were defective in viral systemic movement at the early stage of infection. Interestingly, the reassortant containing TAV RNA1 and CMV RNA2 and RNA3 infected plants systemically, but produced RNA4A (the RNA2 subgenome) at an undetectable level. The defect in production of RNA4A was due to the 1a protein encoded by TAV RNA1, and partially restored by replacing the C-terminus (helicase domain) in TAV 1a with that of CMV 1a. Collectively, exchange of the replicase components between CMV and TAV was acceptable for viral replication, but was defective in either directing transcription of subgenomic RNA4A or facilitating viral long-distance movement. Our finding may shed some light on evolution of subgenomic RNA4A in the family Bromoviridae.

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Huang, M., e S. J. Elledge. "Identification of RNR4, encoding a second essential small subunit of ribonucleotide reductase in Saccharomyces cerevisiae." Molecular and Cellular Biology 17, n.º 10 (outubro de 1997): 6105–13. http://dx.doi.org/10.1128/mcb.17.10.6105.

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Ribonucleotide reductase (RNR), which catalyzes the rate-limiting step for deoxyribonucleotide production required for DNA synthesis, is an alpha2beta2 tetramer consisting of two large and two small subunits. RNR2 encodes a small subunit and is essential for mitotic viability in Saccharomyces cerevisiae. We have cloned a second essential gene encoding a homologous small subunit, RNR4. RNR4 and RNR2 appear to have nonoverlapping functions and cannot substitute for each other even when overproduced. The lethality of RNR4 deletion mutations can be suppressed by overexpression of RNR1 and RNR3, two genes encoding the large subunit of the RNR enzyme, indicating genetic interactions among the RNR genes. RNR2 and RNR4 may be present in the same reductase complex in vivo, since they coimmunoprecipitate from cell extracts. Like the other RNR genes, RNR4 is inducible by DNA-damaging agents through the same signal transduction pathway involving MEC1, RAD53, and DUN1 kinase genes. Analysis of DNA damage inducibility of RNR2 and RNR4 revealed partial inducibility in dun1 mutants, indicating a DUN1-independent branch of the transcriptional response to DNA damage.

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Mohana, K. N., N. Prasad e P. M. Ramadas Bhandarkar. "Determination of Quantum Yield for the Photochemical Decomposition of Dichloramine-B and Dibromamine-B in Aqueous Acetic Acid Medium". E-Journal of Chemistry 4, n.º 4 (2007): 502–9. http://dx.doi.org/10.1155/2007/413836.

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The photolysis of dihaloamines (RNX2),viz., dichloramine-B (DCB) and dibromamine-B (DBB) in aqueous acetic acid (1:1 v/v) solutions has been studied with the UV light source (λ= 2537 Å). The experimental rate law obtained is - d [RNX2] / dt = k' Io/ [RNX2], where Iois the intensity of incident light. The addition of benzenesulphonamide, the product of photolysis or uranyl ion had no significant effect on the rate of photochemical decomposition. A slight decrease in the rate has been observed by the addition of NaCl / NaBr to DCB / DBB solutions. The quantum yield (Φ) for the photolytic decomposition has been computed. A suitable photolytic mechanism and a rate law consistent with the observed results have been proposed.

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