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Single bacterial resolvases first exploit, then constrain intrinsic dynamics of the Holliday junction to direct recombination
Homologous recombination forms and resolves an entangled DNA Holliday Junction (HJ) crucial for achieving genetic reshuffling and genome repair. To maintain genomic integrity, specialized resolvase enzymes cleave the entangled DNA into two discrete DNA molecules. However, it is unclear how two simil...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7969024/ https://www.ncbi.nlm.nih.gov/pubmed/33619520 http://dx.doi.org/10.1093/nar/gkab096 |
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author | Ray, Sujay Pal, Nibedita Walter, Nils G |
author_facet | Ray, Sujay Pal, Nibedita Walter, Nils G |
author_sort | Ray, Sujay |
collection | PubMed |
description | Homologous recombination forms and resolves an entangled DNA Holliday Junction (HJ) crucial for achieving genetic reshuffling and genome repair. To maintain genomic integrity, specialized resolvase enzymes cleave the entangled DNA into two discrete DNA molecules. However, it is unclear how two similar stacking isomers are distinguished, and how a cognate sequence is found and recognized to achieve accurate recombination. We here use single-molecule fluorescence observation and cluster analysis to examine how prototypic bacterial resolvase RuvC singles out two of the four HJ strands and achieves sequence-specific cleavage. We find that RuvC first exploits, then constrains the dynamics of intrinsic HJ isomer exchange at a sampled branch position to direct cleavage toward the catalytically competent HJ conformation and sequence, thus controlling recombination output at minimal energetic cost. Our model of rapid DNA scanning followed by ‘snap-locking’ of a cognate sequence is strikingly consistent with the conformational proofreading of other DNA-modifying enzymes. |
format | Online Article Text |
id | pubmed-7969024 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-79690242021-03-22 Single bacterial resolvases first exploit, then constrain intrinsic dynamics of the Holliday junction to direct recombination Ray, Sujay Pal, Nibedita Walter, Nils G Nucleic Acids Res Nucleic Acid Enzymes Homologous recombination forms and resolves an entangled DNA Holliday Junction (HJ) crucial for achieving genetic reshuffling and genome repair. To maintain genomic integrity, specialized resolvase enzymes cleave the entangled DNA into two discrete DNA molecules. However, it is unclear how two similar stacking isomers are distinguished, and how a cognate sequence is found and recognized to achieve accurate recombination. We here use single-molecule fluorescence observation and cluster analysis to examine how prototypic bacterial resolvase RuvC singles out two of the four HJ strands and achieves sequence-specific cleavage. We find that RuvC first exploits, then constrains the dynamics of intrinsic HJ isomer exchange at a sampled branch position to direct cleavage toward the catalytically competent HJ conformation and sequence, thus controlling recombination output at minimal energetic cost. Our model of rapid DNA scanning followed by ‘snap-locking’ of a cognate sequence is strikingly consistent with the conformational proofreading of other DNA-modifying enzymes. Oxford University Press 2021-02-22 /pmc/articles/PMC7969024/ /pubmed/33619520 http://dx.doi.org/10.1093/nar/gkab096 Text en © The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Nucleic Acid Enzymes Ray, Sujay Pal, Nibedita Walter, Nils G Single bacterial resolvases first exploit, then constrain intrinsic dynamics of the Holliday junction to direct recombination |
title | Single bacterial resolvases first exploit, then constrain intrinsic dynamics of the Holliday junction to direct recombination |
title_full | Single bacterial resolvases first exploit, then constrain intrinsic dynamics of the Holliday junction to direct recombination |
title_fullStr | Single bacterial resolvases first exploit, then constrain intrinsic dynamics of the Holliday junction to direct recombination |
title_full_unstemmed | Single bacterial resolvases first exploit, then constrain intrinsic dynamics of the Holliday junction to direct recombination |
title_short | Single bacterial resolvases first exploit, then constrain intrinsic dynamics of the Holliday junction to direct recombination |
title_sort | single bacterial resolvases first exploit, then constrain intrinsic dynamics of the holliday junction to direct recombination |
topic | Nucleic Acid Enzymes |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7969024/ https://www.ncbi.nlm.nih.gov/pubmed/33619520 http://dx.doi.org/10.1093/nar/gkab096 |
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