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Relevant bibliographies by topics / DNA Polymerase III

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Academic literature on the topic 'DNA Polymerase III'

Author: Grafiati

Published: 4 June 2021

Last updated: 5 March 2023

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Journal articles on the topic "DNA Polymerase III"

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Glover, Bradley P., and Charles S. McHenry. "The DNA Polymerase III Holoenzyme." Cell 105, no. 7 (June 2001): 925–34. http://dx.doi.org/10.1016/s0092-8674(01)00400-7.

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Maslowska, Katarzyna H., Karolina Makiela-Dzbenska, Jin-Yao Mo, Iwona J. Fijalkowska, and Roel M. Schaaper. "High-accuracy lagging-strand DNA replication mediated by DNA polymerase dissociation." Proceedings of the National Academy of Sciences 115, no. 16 (April 2, 2018): 4212–17. http://dx.doi.org/10.1073/pnas.1720353115.

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The fidelity of DNA replication is a critical factor in the rate at which cells incur mutations. Due to the antiparallel orientation of the two chromosomal DNA strands, one strand (leading strand) is replicated in a mostly processive manner, while the other (lagging strand) is synthesized in short sections called Okazaki fragments. A fundamental question that remains to be answered is whether the two strands are copied with the same intrinsic fidelity. In most experimental systems, this question is difficult to answer, as the replication complex contains a different DNA polymerase for each strand, such as, for example, DNA polymerases δ and ε in eukaryotes. Here we have investigated this question in the bacterium Escherichia coli, in which the replicase (DNA polymerase III holoenzyme) contains two copies of the same polymerase (Pol III, the dnaE gene product), and hence the two strands are copied by the same polymerase. Our in vivo mutagenesis data indicate that the two DNA strands are not copied with the same accuracy, and that, remarkably, the lagging strand has the highest fidelity. We postulate that this effect results from the greater dissociative character of the lagging-strand polymerase, which provides additional options for error removal. Our conclusion is strongly supported by results with dnaE antimutator polymerases characterized by increased dissociation rates.

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Budd, M. E., K. D. Wittrup, J. E. Bailey, and J. L. Campbell. "DNA polymerase I is required for premeiotic DNA replication and sporulation but not for X-ray repair in Saccharomyces cerevisiae." Molecular and Cellular Biology 9, no. 2 (February 1989): 365–76. http://dx.doi.org/10.1128/mcb.9.2.365-376.1989.

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We have used a set of seven temperature-sensitive mutants in the DNA polymerase I gene of Saccharomyces cerevisiae to investigate the role of DNA polymerase I in various aspects of DNA synthesis in vivo. Previously, we showed that DNA polymerase I is required for mitotic DNA replication. Here we extend our studies to several stages of meiosis and repair of X-ray-induced damage. We find that sporulation is blocked in all of the DNA polymerase temperature-sensitive mutants and that premeiotic DNA replication does not occur. Commitment to meiotic recombination is only 2% of wild-type levels. Thus, DNA polymerase I is essential for these steps. However, repair of X-ray-induced single-strand breaks is not defective in the DNA polymerase temperature-sensitive mutants, and DNA polymerase I is therefore not essential for repair of such lesions. These results suggest that DNA polymerase II or III or both, the two other nuclear yeast DNA polymerases for which roles have not yet been established, carry out repair in the absence of DNA polymerase I, but that DNA polymerase II and III cannot compensate for loss of DNA polymerase I in meiotic replication and recombination. These results do not, however, rule out essential roles for DNA polymerase II or III or both in addition to that for DNA polymerase I.

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Budd, M. E., K. D. Wittrup, J. E. Bailey, and J. L. Campbell. "DNA polymerase I is required for premeiotic DNA replication and sporulation but not for X-ray repair in Saccharomyces cerevisiae." Molecular and Cellular Biology 9, no. 2 (February 1989): 365–76. http://dx.doi.org/10.1128/mcb.9.2.365.

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We have used a set of seven temperature-sensitive mutants in the DNA polymerase I gene of Saccharomyces cerevisiae to investigate the role of DNA polymerase I in various aspects of DNA synthesis in vivo. Previously, we showed that DNA polymerase I is required for mitotic DNA replication. Here we extend our studies to several stages of meiosis and repair of X-ray-induced damage. We find that sporulation is blocked in all of the DNA polymerase temperature-sensitive mutants and that premeiotic DNA replication does not occur. Commitment to meiotic recombination is only 2% of wild-type levels. Thus, DNA polymerase I is essential for these steps. However, repair of X-ray-induced single-strand breaks is not defective in the DNA polymerase temperature-sensitive mutants, and DNA polymerase I is therefore not essential for repair of such lesions. These results suggest that DNA polymerase II or III or both, the two other nuclear yeast DNA polymerases for which roles have not yet been established, carry out repair in the absence of DNA polymerase I, but that DNA polymerase II and III cannot compensate for loss of DNA polymerase I in meiotic replication and recombination. These results do not, however, rule out essential roles for DNA polymerase II or III or both in addition to that for DNA polymerase I.

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Bonner, C. A., P. T. Stukenberg, M. Rajagopalan, R. Eritja, M. O'Donnell, K. McEntee, H. Echols, and M. F. Goodman. "Processive DNA synthesis by DNA polymerase II mediated by DNA polymerase III accessory proteins." Journal of Biological Chemistry 267, no. 16 (June 1992): 11431–38. http://dx.doi.org/10.1016/s0021-9258(19)49928-6.

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Bryan, Sharon K., Michael Hagensee, and Robb E. Moses. "Holoenzyme DNA polymerase III fixes mutations." Mutation Research Letters 243, no. 4 (April 1990): 313–18. http://dx.doi.org/10.1016/0165-7992(90)90149-e.

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Pritchard, Arthur E., and Charles S. McHenry. "Assembly of DNA Polymerase III Holoenzyme." Journal of Biological Chemistry 276, no. 37 (July 19, 2001): 35217–22. http://dx.doi.org/10.1074/jbc.m102735200.

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Fijalkowska, I. J., and R. M. Schaaper. "Antimutator mutations in the alpha subunit of Escherichia coli DNA polymerase III: identification of the responsible mutations and alignment with other DNA polymerases." Genetics 134, no. 4 (August 1, 1993): 1039–44. http://dx.doi.org/10.1093/genetics/134.4.1039.

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Abstract The dnaE gene of Escherichia coli encodes the DNA polymerase (alpha subunit) of the main replicative enzyme, DNA polymerase III holoenzyme. We have previously identified this gene as the site of a series of seven antimutator mutations that specifically decrease the level of DNA replication errors. Here we report the nucleotide sequence changes in each of the different antimutator dnaE alleles. For each a single, but different, amino acid substitution was found among the 1,160 amino acids of the protein. The observed substitutions are generally nonconservative. All affected residues are located in the central one-third of the protein. Some insight into the function of the regions of polymerase III containing the affected residues was obtained by amino acid alignment with other DNA polymerases. We followed the principles developed in 1990 by M. Delarue et al. who have identified in DNA polymerases from a large number of prokaryotic and eukaryotic sources three highly conserved sequence motifs, which are suggested to contain components of the polymerase active site. We succeeded in finding these three conserved motifs in polymerase III as well. However, none of the amino acid substitutions responsible for the antimutator phenotype occurred at these sites. This and other observations suggest that the effect of these mutations may be exerted indirectly through effects on polymerase conformation and/or DNA/polymerase interactions.

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LOGAN, KELLEY, and STEVEN ACKERMAN. "Effects of Antibiotics on RNA Polymerase III Transcription." DNA 7, no. 7 (September 1988): 483–91. http://dx.doi.org/10.1089/dna.1.1988.7.483.

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De, Ananya, and Colin Campbell. "A novel interaction between DNA ligase III and DNA polymerase γ plays an essential role in mitochondrial DNA stability." Biochemical Journal 402, no. 1 (January 25, 2007): 175–86. http://dx.doi.org/10.1042/bj20061004.

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The data in the present study show that DNA polymerase γ and DNA ligase III interact in mitochondrial protein extracts from cultured HT1080 cells. An interaction was also observed between the two recombinant proteins in vitro. Expression of catalytically inert versions of DNA ligase III that bind DNA polymerase γ was associated with reduced mitochondrial DNA copy number and integrity. In contrast, overexpression of wild-type DNA ligase III had no effect on mitochondrial DNA copy number or integrity. Experiments revealed that wild-type DNA ligase III facilitates the interaction of DNA polymerase γ with a nicked DNA substrate in vitro, and that the zinc finger domain of DNA ligase III is required for this activity. Mitochondrial protein extracts prepared from cells overexpressing a DNA ligase III protein that lacked the zinc finger domain had reduced base excision repair activity compared with extracts from cells overexpressing the wild-type protein. These data support the interpretation that the interaction of DNA ligase III and DNA polymerase γ is required for proper maintenance of the mammalian mitochondrial genome.

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Dissertations / Theses on the topic "DNA Polymerase III"

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Lee, Sally. "Architecture of RNA polymerase II and RNA polymerase III pre-initiation transcription complexes /." Thesis, Connect to this title online; UW restricted, 1997. http://hdl.handle.net/1773/9213.

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Varshney, Dhaval. "Regulation of RNA polymerase III transcription by DNA methylation and chromatin." Thesis, University of Glasgow, 2012. http://theses.gla.ac.uk/3114/.

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Mammalian genomes contain huge numbers of short interspersed elements (SINEs). An extreme case is provided by the human genome, which carries ~106 copies of Alu SINEs that together account for ~10% of total chromosomal DNA. SINEs spread by retrotransposition, which depends on their transcription by pol III. This transcription is heavily suppressed. Silencing is thought to involve DNA methylation and packaging the SINEs into chromatin structures that deny access of transcription factors. It has been argued that this may be of great importance to prevent SINEs from competing with essential genes for a limited pool of transcription machinery. Our investigation of this has revealed some unexpected findings. This study has also investigated the effects of SWI/SNF chromatin remodellers on tRNA transcription.

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Allison, Simon J. "Regulatory studies of the mammalian RNA polymerase III transcriptional apparatus." Thesis, University of Glasgow, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.343976.

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Isoz, Isabelle. "Role of yeast DNA polymerase epsilon during DNA replication." Doctoral thesis, Umeå : Umeå University, 2008. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-1932.

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Slater, Steven Charles. "The role of DNA polymerase III in DNA repair and mutagenesis in Escherichia coli and Salmonella typhimurium." Case Western Reserve University School of Graduate Studies / OhioLINK, 1994. http://rave.ohiolink.edu/etdc/view?acc_num=case1057261938.

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Colbert, Trenton. "Characterization of BRF1, an RNA polymerase III transcription factor /." Thesis, Connect to this title online; UW restricted, 1997. http://hdl.handle.net/1773/6320.

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Pacitti, Diane Frances. "The Characterization of Staphylococcus Aureus polC: the Structural Gene for DNA Polymerase III." eScholarship@UMMS, 1995. http://escholarship.umassmed.edu/gsbs_diss/271.

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The major research interest of our laboratory is focused on the replication-specific DNA polymerase III (pol III) family in Gram+ bacteria, and has used Bacillus subtilis (BS) as the primary model enzyme for study. The long range objective of the work of the laboratory is to gain a deeper understanding of the structure and function of Gram+ bacterial DNA polymerase IIIs, a structurally unique class of DNA-dependent DNA polymerase which are uniquely susceptible to inhibition by a specific class of dGTP analogs. The project described in this thesis dissertation deals specifically with the pol III of the Gram+ organism Staphylococcus aureus, and involves the isolation and characterization of DNA pol III from this clinically relevant pathogenic bacterium. A homology-based strategy was devised to clone the structural gene specifying DNA polymerase III of Staphylococcus aureus, SA polC. SA polC was found to contain a 4305-bp open reading frame (ORF) encoding a 162.4 kDa polypeptide, and mapped between Ω1074[Tn551] and recA/ngr on the genome map of S. aureus NCTC 8325. The 1435 codon ORF was engineered into the E. coli expression plasmid pBS(KS) under the control of the lac promoter and its repressor. The translational signals of SA polC were reengineered using expression cassette PCR (ECPCR) to optimize the in vitro expression of SA polC in E. coli. Derepression of E. coli transformants carrying the recombinant vector generated high level expression of active recombinant pol III. The recombinant SA pol III was purified to greater than 98% homogeneity and was shown by N-terminal amino acid analysis to be the bona fide product of the 4305-bp SA polC ORF. The physical and catalytic properties of recombinant SA pol III and its responsiveness to inhibitors of the HPUra type were similar to those of Bacillus subtilis (BS) pol III. Comparative structural analysis of the primary structure of SA pol III and the pol IIIs of B. subtilis and the Gram+ relative Mycoplasma pulmonis indicated strong conservation of essential catalytic domains and a novel zinc-finger motif. Comparison of the primary structures of E. coli pol III and these three Gram+ enzymes suggested a specific evolutionary relationship between the pol IIIs of Gram+ and Gram- bacteria.

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Lancy, Edward Donald Jr. "Genetic requirements for growth of Salmonella typhimurium lacking the proofreading subunit of DNA polymerase III." Case Western Reserve University School of Graduate Studies / OhioLINK, 1990. http://rave.ohiolink.edu/etdc/view?acc_num=case1054846339.

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Sabouri, Nasim. "Structure of eukaryotic DNA polymerase epsilon and lesion bypass capability." Doctoral thesis, Umeå : Univ, 2008. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-1477.

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Hög, Friederike. "Functional studies of RNA polymerase II recruitment to promoter DNA and impact of BRF1 mutations on RNA polymerase III-dependent transcription." Diss., Ludwig-Maximilians-Universität München, 2014. http://nbn-resolving.de/urn:nbn:de:bvb:19-179326.

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Books on the topic "DNA Polymerase III"

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(Editor), Sankar Adhya, and Susan Garges (Editor), eds. RNA Polymerase and Associated Factors, Part C, Volume 370 (Methods in Enzymology). Academic Press, 2003.

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Book chapters on the topic "DNA Polymerase III"

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McHenry, Charles. "DNA Polymerase III Structure." In Molecular Life Sciences, 1–10. New York, NY: Springer New York, 2014. http://dx.doi.org/10.1007/978-1-4614-6436-5_131-1.

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McHenry, Charles S. "DNA Polymerase III Structure." In Molecular Life Sciences, 210–17. New York, NY: Springer New York, 2018. http://dx.doi.org/10.1007/978-1-4614-1531-2_131.

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Furth, John J. "RNA Polymerase III and Transcription of 5S Ribosomal DNA." In Molecular Biology of Chromosome Function, 207–23. New York, NY: Springer New York, 1989. http://dx.doi.org/10.1007/978-1-4612-3652-8_9.

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Merkl, Philipp E., Christopher Schächner, Michael Pilsl, Katrin Schwank, Catharina Schmid, Gernot Längst, Philipp Milkereit, Joachim Griesenbeck, and Herbert Tschochner. "Specialization of RNA Polymerase I in Comparison to Other Nuclear RNA Polymerases of Saccharomyces cerevisiae." In Ribosome Biogenesis, 63–70. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-2501-9_4.

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AbstractIn archaea and bacteria the major classes of RNAs are synthesized by one DNA-dependent RNA polymerase (RNAP). In contrast, most eukaryotes have three highly specialized RNAPs to transcribe the nuclear genome. RNAP I synthesizes almost exclusively ribosomal (r)RNA, RNAP II synthesizes mRNA as well as many noncoding RNAs involved in RNA processing or RNA silencing pathways and RNAP III synthesizes mainly tRNA and 5S rRNA. This review discusses functional differences of the three nuclear core RNAPs in the yeast S. cerevisiae with a particular focus on RNAP I transcription of nucleolar ribosomal (r)DNA chromatin.

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O’donnell, M., J. Kuriyan, X. P. Kong, P. T. Stukenberg, R. Onrust, and N. Yao. "The β Sliding Clamp of E. coli DNA Polymerase III Holoenzyme Balances Opposing Functions." In Nucleic Acids and Molecular Biology, 197–216. Berlin, Heidelberg: Springer Berlin Heidelberg, 1994. http://dx.doi.org/10.1007/978-3-642-78666-2_11.

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Bridges, B. A. "Ultraviolet Light Mutagenesis in Bacteria: The Possible Role of a DNA Polymerase III Complex Lacking Proofreading Exonuclease." In Mechanisms of Environmental Mutagenesis-Carcinogenesis, 27–35. Boston, MA: Springer US, 1990. http://dx.doi.org/10.1007/978-1-4615-3808-0_2.

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Merkl, Philipp E., Christopher Schächner, Michael Pilsl, Katrin Schwank, Kristin Hergert, Gernot Längst, Philipp Milkereit, Joachim Griesenbeck, and Herbert Tschochner. "Analysis of Yeast RNAP I Transcription of Nucleosomal Templates In Vitro." In Ribosome Biogenesis, 39–59. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-2501-9_3.

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AbstractNuclear eukaryotic RNA polymerases (RNAPs) transcribe a chromatin template in vivo. Since the basic unit of chromatin, the nucleosome, renders the DNA largely inaccessible, RNAPs have to overcome the nucleosomal barrier for efficient RNA synthesis. Gaining mechanistical insights in the transcription of chromatin templates will be essential to understand the complex process of eukaryotic gene expression. In this article we describe the use of defined in vitro transcription systems for comparative analysis of highly purified RNAPs I–III from S. cerevisiae (hereafter called yeast) transcribing in vitro reconstituted nucleosomal templates. We also provide a protocol to study promoter-dependent RNAP I transcription of purified native 35S ribosomal RNA (rRNA) gene chromatin.

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Butler, Michelle M., and George E. Wright. "A Method to Assay Inhibitors of DNA Polymerase IIIC Activity." In Methods In Molecular Medicine™, 25–36. Totowa, NJ: Humana Press, 2008. http://dx.doi.org/10.1007/978-1-59745-246-5_3.

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Thomas, David C., John D. Roberts, Mary P. Fitzgerald, and Thomas A. Kunkel. "Fidelity of Animal Cell DNA Polymerases α and δ and of a Human DNA Replication Complex." In Antimutagenesis and Anticarcinogenesis Mechanisms II, 289–97. Boston, MA: Springer US, 1990. http://dx.doi.org/10.1007/978-1-4615-9561-8_24.

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Reuter, C., Ulrike Auf der Landwehr, M. Zühlsdorf, C. Busemann, B. Wörmann, T. Büchner, and W. Hiddemann. "Recombinant Human Granulocyte-Macrophage Colony-Stimulating Factor Increases DNA Polymerase Activity in Acute Myeloid Leukemia Blasts In Vitro and In Vivo." In Cytokines in Hemopoiesis, Oncology, and AIDS II, 321–28. Berlin, Heidelberg: Springer Berlin Heidelberg, 1992. http://dx.doi.org/10.1007/978-3-642-48715-6_41.

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Conference papers on the topic "DNA Polymerase III"

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SILVEIRA, ANA CAROLINA ATAIDE, JEFFERSON JOE MOREIRA ALVES, LUAN SOUZA DE PAULA GOMES, MATHEUS HENRIQUE TEIXEIRA, and DEMERSON ARRUDA SANGLARD. "VALIDAÇÃO DE UM PROTOCOLO ÁGIL E EFICAZ PARA EXTRAÇÃO DE DNA DE FOLHAS MADURAS DE ACESSOS DE MILHOS CRIOULOS." In III Congresso Brasileiro de Ciências Biologicas. Revista Multidisciplinar de Educação e Meio Ambiente, 2022. http://dx.doi.org/10.51189/iii-conbracib/7280.

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Introdução: A extração de DNA é um dos principais procedimentos para a utilização de técnicas moleculares. Possui enorme importância nos protocolos de amplificação via Reação em Cadeia da Polimerase (Polymerase Chain Reaction - PCR). Esta, por sua vez, permite estimar a dissimilaridade genética entre acessos de uma população, selecionar genes específicos, realizar estudos de alelismo, construir mapas genéticos, obter clonagens posicionais, dentre muitos outros procedimentos úteis ao melhoramento genético.Objetivos: Validar um protocolo de extração de DNA com rapidez e elevada pureza, a partir de folhas maduras de 44 acessos de milhos crioulos prospectados no Norte de Minas Gerais. (Zea mays L.).Material e métodos: Testou-se procedimentos modificados baseados no protocolo de base CTAB (Brometo de Cetil Trimetilamonio ou Cetyl trimethylammonium bromid). Nesse protocolo foram utilizadas diferentes concentrações de β-mercaptoetanol no tampão de extração (0,0; 0,2; 10; 15; 25; e 50 uL de β-mercaptoetanol/mL), considerando o seguinte tampão de extração: Tris-HCl 100 mM em pH 8; EDTA 20 mM; NaCl 1,4 mM; CTAB 2% e; PVP 1%). Os procedimentos foram conduzidos no Laboratório de Biotecnologia da Universidade Federal de Minas Gerais (campus Montes Claros), fomentados pelo Banco do Nordeste (Fundo de Desenvolvimento Econômico, Científico, Tecnológico e de Inovação - FUNDECI).Resultados: O protocolo foi eficiente no isolamento de DNA livre de polissacarídeos e polifenóis, com rendimento do DNA de alto peso molecular e concentrações acima de 120 ng/uL, utilizando-se valores superiores a 1% de β-mercaptoetanol no tampão de extração. Outras modificações importantes se ativeram aos passos da maceração, substituindo o uso do nitrogênio líquido (trituração das amostras em cadinho e pistilo), por uma ação direta do aparelho disruptor/homogeneizador de amostras biológicas, nos microtubos acrescidos do tampão. Além disso, adicionou-se duas rodadas para desproteinização (clorofórmio álcool isomílico 24:1), sendo os dois passos de pousios resfriados das amostras realizados apenas em ultrafreezer (-75oC) durante 10 min (cerca de 48 h poupadas).Conclusão: O protocolo de base CTAB para extração de DNA, inicialmente proposto para folhas de plantas leguminosas (largas), também foi eficaz em folhas maduras de acessos de milhos crioulos (gramíneas), levando em conta adaptações que permitiram pureza e celeridade ao processo.

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Ramos, Amanda Bruno da Silva Bellini, Gabriela de Souza Martins Faria, Tayna Aparecida Marques de Carvalho, Fábio Antônio Colombo, and Angélica Rosa. "MONITORAMENTO DA TRANSMISSÃO VETORIAL DE TRYPANOSOMA CRUZI NO SUL DE MINAS GERAIS." In II Congresso Brasileiro de Parasitologia Humana On-line. Revista Multidisciplinar em Saúde, 2022. http://dx.doi.org/10.51161/conbrapah/3.

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Introdução: A Doença de Chagas, cujo agente etiológico é o Trypanosoma cruzi, ainda é problema de saúde pública em muitos estados do Brasil. Por se tratar de uma enfermidade de transmissão vetorial, a maioria das formas de monitoramento da mesma perpassa pelo estudo do vetor. Dessa forma, a Universidade Federal de Alfenas, em parceria com a Superintendência Regional de Saúde/Alfenas, tem desenvolvido o trabalho de vigilância epidemiológica da Doença de Chagas na região, estudo indispensável considerando os diversos fatores de risco aqui existentes. Objetivo: Assim sendo, o objetivo desse trabalho é: Identificar os insetos e o agente etiológico da Doença de Chagas em fezes de triatomíneos capturados pela população e/ou agentes de saúde dos municípios sob a jurisdição da Vigilância Epidemiológica da SRS/Alfenas, além de emitir laudos baseados nos diagnósticos parasitológico e molecular e realizar comparação com estudos anteriores. Material e Métodos: A análise entomológica é feita de acordo com características morfológicas dos insetos; para análise parasitológica, observa-se as fezes dos insetos em lâmina à fresco, em microscópio óptico. Por fim, a análise molecular é feita pela técnica Real Time Polymerase Chain Reaction (qPCR), com DNA extraído das fezes. Resultados: Entre Julho de 2020 e Junho de 2021, foram recebidos 209 insetos: 95,21% identificados como triatomíneos, e 4,79% como outros hemípteros (fitófagos e predadores). Dentre os triatomíneos, 137 foram recebidos na segunda metade de 2020: 2 identificados como Rhodnius neglectus e 135 como Panstrongylus megistus. As fezes dos insetos foram analisadas por exame parasitológico direto a fresco, com 10,95% de positividade para flagelados semelhantes ao T. cruzi, e também pela técnica molecular (qPCR), que detectou DNA de T. cruzi em 20,44% das amostras. Os demais 62 triatomíneos foram recebidos no primeiro semestre de 2021, sendo todos identificados como Panstrongylus megistus. Destes, 22,58% testaram positivo na análise parasitológica, e 37,10% na análise molecular. Em comparação coma média dos últimos 7 anos (2014- 2020), a porcentagem de amostras positivas pelo diagnóstico molecular aumentou em 10% na primeira metade de 2021, o que indica que o trabalho de vigilância deve continuar e os cuidados com relação a essa parasitose devem receber atenção.

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Vieira, Paula Cristina de Vasconcelos, Adriano de Paula Sabino, Jaqueline Germano de Oliveira, Marcelo Antônio Pascoal Xavier, Annamaria Ravara Vago, and Maria Gabrielle de Lima Rocha. "Avaliação de RAP1 como biomarcador da neoplasia intraepitelial cervical." In XIII Congresso da Sociedade Brasileira de DST - IX Congresso Brasileiro de AIDS - IV Congresso Latino Americano de IST/HIV/AIDS. Zeppelini Editorial e Comunicação, 2021. http://dx.doi.org/10.5327/dst-2177-8264-202133p067.

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Introdução: Mundialmente, o carcinoma de colo uterino associado ao papilomavírus humano está entre as maiores causas de morte relacionadas ao câncer na população feminina. Nos últimos anos, a detecção de lesões pré-neoplásicas mediante a ação de programas consolidados de rastreamento, prevenção e acompanhamento clínico permitiram a redução das taxas de câncer de colo em diversos países. Entretanto, essa neoplasia ainda é muito frequente. Esse problema tem fortalecido a procura por biomarcadores que possam aumentar a eficiência do rastreamento. Entre esses biomarcadores está RAP1, pequena GTPase com atuação em múltiplas vias de sinalização celular e em processos neoplásicos. Objetivo: Avaliar, em biópsias de colo uterino diagnosticadas por histopatologia em cervicites, lesões pré-neoplásicas e neoplásicas, (i) a expressão de RAP1 por imuno-histoquímica nas áreas lesionadas do epitélio, (ii) a prevalência da infecção por papilomavírus humano por meio da Nested Polymerase Chain Reaction e a genotipagem dos papilomavírus humano por sequenciamento automático de ácido desoxirribonucleico. Resultados: A presença do ácido desozirribonucleico do vírus da imunodeficiência humana foi observada em 74% das amostras, e o genótipo do vírus da imunodeficiência humana 16 foi o mais prevalente, especialmente em neoplasia intraepitelial cervical grau III e câncer. Em relação a RAP1 em tecidos cervicais, observou-se a expressão da proteína em amostras de todos os grupos analisados. A marcação esteve predominantemente no núcleo e no citoplasma de células epiteliais, representando 69% das amostras cervicais. Em relação ao parâmetro intensidade da marcação, verificou-se que amostras de neoplasia intraepitelial cervical grau III exibiram maior intensidade de RAP1 do que lesões de menor gravidade (Cervicites + neoplasia intraepitelial cervical grau I). Nas amostras de carcinoma de células escamosas também se observou a imunomarcação para RAP1 predominantemente intensa, com forte marcação nuclear. Percebe-se que nas lesões mais graves, neoplasia intraepitelial cervical grau III e carcinoma de células escamosas a quantidade de núcleos marcados é mais elevada se comparado a lesões mais leves. Conclusão: Uso potencial da marcação tecidual de RAP1 como biomarcador de lesões cervicais de maior gravidade, como neoplasia intraepitelial cervical grau III e carcinoma de células escamosas.

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Reports on the topic "DNA Polymerase III"

1

Laurence, Jeffrey. Antibody to the RNA-Dependent DNA Polymerase of HTLV-III: Characterization and Clinical Associations. Fort Belvoir, VA: Defense Technical Information Center, March 1990. http://dx.doi.org/10.21236/ada231466.

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2

Laurence, Jeffrey. Antibody to the RNA-Dependent DNA Polymerase of HTLV-III: characterization and Clinical Associations. Fort Belvoir, VA: Defense Technical Information Center, July 1988. http://dx.doi.org/10.21236/ada227404.

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3

Laurence, Jeffrey. Antibody to the RNA-Dependent DNA Polymerase of HTLV-III: characterization and Clinical Associations. Fort Belvoir, VA: Defense Technical Information Center, December 1988. http://dx.doi.org/10.21236/ada227519.

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4

Altstein, Miriam, and Ronald J. Nachman. Rational Design of Insect Control Agent Prototypes Based on Pyrokinin/PBAN Neuropeptide Antagonists. United States Department of Agriculture, August 2013. http://dx.doi.org/10.32747/2013.7593398.bard.

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The general objective of this study was to develop rationally designed mimetic antagonists (and agonists) of the PK/PBAN Np class with enhanced bio-stability and bioavailability as prototypes for effective and environmentally friendly pest insect management agents. The PK/PBAN family is a multifunctional group of Nps that mediates key functions in insects (sex pheromone biosynthesis, cuticular melanization, myotropic activity, diapause and pupal development) and is, therefore, of high scientific and applied interest. The objectives of the current study were: (i) to identify an antagonist biophores (ii) to develop an arsenal of amphiphilic topically active PK/PBAN antagonists with an array of different time-release profiles based on the previously developed prototype analog; (iii) to develop rationally designed non-peptide SMLs based on the antagonist biophore determined in (i) and evaluate them in cloned receptor microplate binding assays and by pheromonotropic, melanotropic and pupariation in vivo assays. (iv) to clone PK/PBAN receptors (PK/PBAN-Rs) for further understanding of receptor-ligand interactions; (v) to develop microplate binding assays for screening the above SMLs. In the course of the granting period A series of amphiphilic PK/PBAN analogs based on a linear lead antagonist from the previous BARD grant was synthesized that incorporated a diverse array of hydrophobic groups (HR-Suc-A[dF]PRLa). Others were synthesized via the attachment of polyethylene glycol (PEG) polymers. A hydrophobic, biostablePK/PBAN/DH analog DH-2Abf-K prevented the onset of the protective state of diapause in H. zea pupae [EC50=7 pmol/larva] following injection into the preceding larval stage. It effectively induces the crop pest to commit a form of ‘ecological suicide’. Evaluation of a set of amphiphilic PK analogs with a diverse array of hydrophobic groups of the formula HR-Suc-FTPRLa led to the identification of analog T-63 (HR=Decyl) that increased the extent of diapause termination by a factor of 70% when applied topically to newly emerged pupae. Another biostablePK analog PK-Oic-1 featured anti-feedant and aphicidal properties that matched the potency of some commercial aphicides. Native PK showed no significant activity. The aphicidal effects were blocked by a new PEGylated PK antagonist analog PK-dF-PEG4, suggesting that the activity is mediated by a PK/PBAN receptor and therefore indicative of a novel and selective mode-of-action. Using a novel transPro mimetic motif (dihydroimidazole; ‘Jones’) developed in previous BARD-sponsored work, the first antagonist for the diapause hormone (DH), DH-Jo, was developed and shown to block over 50% of H. zea pupal diapause termination activity of native DH. This novel antagonist development strategy may be applicable to other invertebrate and vertebrate hormones that feature a transPro in the active core. The research identifies a critical component of the antagonist biophore for this PK/PBAN receptor subtype, i.e. a trans-oriented Pro. Additional work led to the molecular cloning and functional characterization of the DH receptor from H. zea, allowing for the discovery of three other DH antagonist analogs: Drosophila ETH, a β-AA analog, and a dF analog. The receptor experiments identified an agonist (DH-2Abf-dA) with a maximal response greater than native DH. ‘Deconvolution’ of a rationally-designed nonpeptide heterocyclic combinatorial library with a cyclic bis-guanidino (BG) scaffold led to discovery of several members that elicited activity in a pupariation acceleration assay, and one that also showed activity in an H. zea diapause termination assay, eliciting a maximal response of 90%. Molecular cloning and functional characterization of a CAP2b antidiuretic receptor from the kissing bug (R. prolixus) as well as the first CAP2b and PK receptors from a tick was also achieved. Notably, the PK/PBAN-like receptor from the cattle fever tick is unique among known PK/PBAN and CAP2b receptors in that it can interact with both ligand types, providing further evidence for an evolutionary relationship between these two NP families. In the course of the granting period we also managed to clone the PK/PBAN-R of H. peltigera, to express it and the S. littoralis-R Sf-9 cells and to evaluate their interaction with a variety of PK/PBAN ligands. In addition, three functional microplate assays in a HTS format have been developed: a cell-membrane competitive ligand binding assay; a Ca flux assay and a whole cell cAMP ELISA. The Ca flux assay has been used for receptor characterization due to its extremely high sensitivity. Computer homology studies were carried out to predict both receptor’s SAR and based on this analysis 8 mutants have been generated. The bioavailability of small linear antagonistic peptides has been evaluated and was found to be highly effective as sex pheromone biosynthesis inhibitors. The activity of 11 new amphiphilic analogs has also been evaluated. Unfortunately, due to a problem with the Heliothis moth colony we were unable to select those with pheromonotropic antagonistic activity and further check their bioavailability. Six peptides exhibited some melanotropic antagonistic activity but due to the low inhibitory effect the peptides were not further tested for bioavailability in S. littoralis larvae. Despite the fact that no new antagonistic peptides were discovered in the course of this granting period the results contribute to a better understanding of the interaction of the PK/PBAN family of Nps with their receptors, provided several HT assays for screening of libraries of various origin for presence of PK/PBAN-Ragonists and antagonists and provided important practical information for the further design of new, peptide-based insecticide prototypes aimed at the disruption of key neuroendocrine physiological functions in pest insects.

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