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Molecular mechanism of sequence-dependent stability of RecA filament

RecA is a DNA-dependent ATPase and mediates homologous recombination by first forming a filament on a single-stranded (ss) DNA. RecA binds preferentially to TGG repeat sequence, which resembles the recombination hot spot Chi (5′-GCTGGTGG-3′) and is the most frequent pattern (GTG) of the codon usage...

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Autores principales: Kim, Sung Hyun, Joo, Chirlmin, Ha, Taekjip, Kim, Doseok
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3763553/
https://www.ncbi.nlm.nih.gov/pubmed/23804763
http://dx.doi.org/10.1093/nar/gkt570
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author Kim, Sung Hyun
Joo, Chirlmin
Ha, Taekjip
Kim, Doseok
author_facet Kim, Sung Hyun
Joo, Chirlmin
Ha, Taekjip
Kim, Doseok
author_sort Kim, Sung Hyun
collection PubMed
description RecA is a DNA-dependent ATPase and mediates homologous recombination by first forming a filament on a single-stranded (ss) DNA. RecA binds preferentially to TGG repeat sequence, which resembles the recombination hot spot Chi (5′-GCTGGTGG-3′) and is the most frequent pattern (GTG) of the codon usage in Escherichia coli. Because of the highly dynamic nature of RecA filament formation, which consists of filament nucleation, growth and shrinkage, we need experimental approaches that can resolve each of these processes separately to gain detailed insights into the molecular mechanism of sequence preference. By using a single-molecule fluorescence assay, we examined the effect of sequence on individual stages of nucleation, monomer binding and dissociation. We found that RecA does not recognize the Chi sequence as a nucleation site. In contrast, we observed that it is the reduced monomer dissociation that mainly determines the high filament stability on TGG repeats. This sequence dependence of monomer dissociation is well-correlated with that of ATP hydrolysis, suggesting that DNA sequence dictates filament stability through modulation of ATP hydrolysis.
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spelling pubmed-37635532013-09-10 Molecular mechanism of sequence-dependent stability of RecA filament Kim, Sung Hyun Joo, Chirlmin Ha, Taekjip Kim, Doseok Nucleic Acids Res Genome Integrity, Repair and Replication RecA is a DNA-dependent ATPase and mediates homologous recombination by first forming a filament on a single-stranded (ss) DNA. RecA binds preferentially to TGG repeat sequence, which resembles the recombination hot spot Chi (5′-GCTGGTGG-3′) and is the most frequent pattern (GTG) of the codon usage in Escherichia coli. Because of the highly dynamic nature of RecA filament formation, which consists of filament nucleation, growth and shrinkage, we need experimental approaches that can resolve each of these processes separately to gain detailed insights into the molecular mechanism of sequence preference. By using a single-molecule fluorescence assay, we examined the effect of sequence on individual stages of nucleation, monomer binding and dissociation. We found that RecA does not recognize the Chi sequence as a nucleation site. In contrast, we observed that it is the reduced monomer dissociation that mainly determines the high filament stability on TGG repeats. This sequence dependence of monomer dissociation is well-correlated with that of ATP hydrolysis, suggesting that DNA sequence dictates filament stability through modulation of ATP hydrolysis. Oxford University Press 2013-09 2013-06-26 /pmc/articles/PMC3763553/ /pubmed/23804763 http://dx.doi.org/10.1093/nar/gkt570 Text en © The Author(s) 2013. Published by Oxford University Press. http://creativecommons.org/licenses/by/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Genome Integrity, Repair and Replication
Kim, Sung Hyun
Joo, Chirlmin
Ha, Taekjip
Kim, Doseok
Molecular mechanism of sequence-dependent stability of RecA filament
title Molecular mechanism of sequence-dependent stability of RecA filament
title_full Molecular mechanism of sequence-dependent stability of RecA filament
title_fullStr Molecular mechanism of sequence-dependent stability of RecA filament
title_full_unstemmed Molecular mechanism of sequence-dependent stability of RecA filament
title_short Molecular mechanism of sequence-dependent stability of RecA filament
title_sort molecular mechanism of sequence-dependent stability of reca filament
topic Genome Integrity, Repair and Replication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3763553/
https://www.ncbi.nlm.nih.gov/pubmed/23804763
http://dx.doi.org/10.1093/nar/gkt570
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