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Human Rev1 polymerase disrupts G-quadruplex DNA

The Y-family DNA polymerase Rev1 is required for successful replication of G-quadruplex DNA (G4 DNA) in higher eukaryotes. Here we show that human Rev1 (hRev1) disrupts G4 DNA structures and prevents refolding in vitro. Nucleotidyl transfer by hRev1 is not necessary for mechanical unfolding to occur...

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Autores principales: Eddy, Sarah, Ketkar, Amit, Zafar, Maroof K., Maddukuri, Leena, Choi, Jeong-Yun, Eoff, Robert L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2014
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3950705/
https://www.ncbi.nlm.nih.gov/pubmed/24366879
http://dx.doi.org/10.1093/nar/gkt1314
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author Eddy, Sarah
Ketkar, Amit
Zafar, Maroof K.
Maddukuri, Leena
Choi, Jeong-Yun
Eoff, Robert L.
author_facet Eddy, Sarah
Ketkar, Amit
Zafar, Maroof K.
Maddukuri, Leena
Choi, Jeong-Yun
Eoff, Robert L.
author_sort Eddy, Sarah
collection PubMed
description The Y-family DNA polymerase Rev1 is required for successful replication of G-quadruplex DNA (G4 DNA) in higher eukaryotes. Here we show that human Rev1 (hRev1) disrupts G4 DNA structures and prevents refolding in vitro. Nucleotidyl transfer by hRev1 is not necessary for mechanical unfolding to occur. hRev1 binds G4 DNA substrates with K(d,DNA) values that are 4–15-fold lower than those of non-G4 DNA substrates. The pre-steady-state rate constant of deoxycytidine monophosphate (dCMP) insertion opposite the first tetrad-guanine by hRev1 is ∼56% as fast as that observed for non-G4 DNA substrates. Thus, hRev1 can promote fork progression by either dislodging tetrad guanines to unfold the G4 DNA, which could assist in extension by other DNA polymerases, or hRev1 can prevent refolding of G4 DNA structures. The hRev1 mechanism of action against G-quadruplexes helps explain why replication progress is impeded at G4 DNA sites in Rev1-deficient cells and illustrates another unique feature of this enzyme with important implications for genome maintenance.
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spelling pubmed-39507052014-03-12 Human Rev1 polymerase disrupts G-quadruplex DNA Eddy, Sarah Ketkar, Amit Zafar, Maroof K. Maddukuri, Leena Choi, Jeong-Yun Eoff, Robert L. Nucleic Acids Res The Y-family DNA polymerase Rev1 is required for successful replication of G-quadruplex DNA (G4 DNA) in higher eukaryotes. Here we show that human Rev1 (hRev1) disrupts G4 DNA structures and prevents refolding in vitro. Nucleotidyl transfer by hRev1 is not necessary for mechanical unfolding to occur. hRev1 binds G4 DNA substrates with K(d,DNA) values that are 4–15-fold lower than those of non-G4 DNA substrates. The pre-steady-state rate constant of deoxycytidine monophosphate (dCMP) insertion opposite the first tetrad-guanine by hRev1 is ∼56% as fast as that observed for non-G4 DNA substrates. Thus, hRev1 can promote fork progression by either dislodging tetrad guanines to unfold the G4 DNA, which could assist in extension by other DNA polymerases, or hRev1 can prevent refolding of G4 DNA structures. The hRev1 mechanism of action against G-quadruplexes helps explain why replication progress is impeded at G4 DNA sites in Rev1-deficient cells and illustrates another unique feature of this enzyme with important implications for genome maintenance. Oxford University Press 2014-03 2013-12-22 /pmc/articles/PMC3950705/ /pubmed/24366879 http://dx.doi.org/10.1093/nar/gkt1314 Text en © The Author(s) 2013. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), 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 Eddy, Sarah
Ketkar, Amit
Zafar, Maroof K.
Maddukuri, Leena
Choi, Jeong-Yun
Eoff, Robert L.
Human Rev1 polymerase disrupts G-quadruplex DNA
title Human Rev1 polymerase disrupts G-quadruplex DNA
title_full Human Rev1 polymerase disrupts G-quadruplex DNA
title_fullStr Human Rev1 polymerase disrupts G-quadruplex DNA
title_full_unstemmed Human Rev1 polymerase disrupts G-quadruplex DNA
title_short Human Rev1 polymerase disrupts G-quadruplex DNA
title_sort human rev1 polymerase disrupts g-quadruplex dna
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3950705/
https://www.ncbi.nlm.nih.gov/pubmed/24366879
http://dx.doi.org/10.1093/nar/gkt1314
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