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Concerted and differential actions of two enzymatic domains underlie Rad5 contributions to DNA damage tolerance
Many genome maintenance factors have multiple enzymatic activities. In most cases, how their distinct activities functionally relate with each other is unclear. Here we examined the conserved budding yeast Rad5 protein that has both ubiquitin ligase and DNA helicase activities. The Rad5 ubiquitin li...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4357696/ https://www.ncbi.nlm.nih.gov/pubmed/25690888 http://dx.doi.org/10.1093/nar/gkv004 |
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author | Choi, Koyi Batke, Sabrina Szakal, Barnabas Lowther, Jonathan Hao, Fanfan Sarangi, Prabha Branzei, Dana Ulrich, Helle D. Zhao, Xiaolan |
author_facet | Choi, Koyi Batke, Sabrina Szakal, Barnabas Lowther, Jonathan Hao, Fanfan Sarangi, Prabha Branzei, Dana Ulrich, Helle D. Zhao, Xiaolan |
author_sort | Choi, Koyi |
collection | PubMed |
description | Many genome maintenance factors have multiple enzymatic activities. In most cases, how their distinct activities functionally relate with each other is unclear. Here we examined the conserved budding yeast Rad5 protein that has both ubiquitin ligase and DNA helicase activities. The Rad5 ubiquitin ligase activity mediates PCNA poly-ubiquitination and subsequently recombination-based DNA lesion tolerance. Interestingly, the ligase domain is embedded in a larger helicase domain comprising seven consensus motifs. How features of the helicase domain influence ligase function is controversial. To clarify this issue, we use genetic, 2D gel and biochemical analyses and show that a Rad5 helicase motif important for ATP binding is also required for PCNA poly-ubiquitination and recombination-based lesion tolerance. We determine that this requirement is due to a previously unrecognized contribution of the motif to the PCNA and ubiquitination enzyme interaction, and not due to its canonical role in supporting helicase activity. We further show that Rad5′s helicase-mediated contribution to replication stress survival is separable from recombination. These findings delineate how two Rad5 enzymatic domains concertedly influence PCNA modification, and unveil their discrete contributions to stress tolerance. |
format | Online Article Text |
id | pubmed-4357696 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-43576962015-03-20 Concerted and differential actions of two enzymatic domains underlie Rad5 contributions to DNA damage tolerance Choi, Koyi Batke, Sabrina Szakal, Barnabas Lowther, Jonathan Hao, Fanfan Sarangi, Prabha Branzei, Dana Ulrich, Helle D. Zhao, Xiaolan Nucleic Acids Res Genome Integrity, Repair and Replication Many genome maintenance factors have multiple enzymatic activities. In most cases, how their distinct activities functionally relate with each other is unclear. Here we examined the conserved budding yeast Rad5 protein that has both ubiquitin ligase and DNA helicase activities. The Rad5 ubiquitin ligase activity mediates PCNA poly-ubiquitination and subsequently recombination-based DNA lesion tolerance. Interestingly, the ligase domain is embedded in a larger helicase domain comprising seven consensus motifs. How features of the helicase domain influence ligase function is controversial. To clarify this issue, we use genetic, 2D gel and biochemical analyses and show that a Rad5 helicase motif important for ATP binding is also required for PCNA poly-ubiquitination and recombination-based lesion tolerance. We determine that this requirement is due to a previously unrecognized contribution of the motif to the PCNA and ubiquitination enzyme interaction, and not due to its canonical role in supporting helicase activity. We further show that Rad5′s helicase-mediated contribution to replication stress survival is separable from recombination. These findings delineate how two Rad5 enzymatic domains concertedly influence PCNA modification, and unveil their discrete contributions to stress tolerance. Oxford University Press 2015-03-11 2015-02-17 /pmc/articles/PMC4357696/ /pubmed/25690888 http://dx.doi.org/10.1093/nar/gkv004 Text en © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Genome Integrity, Repair and Replication Choi, Koyi Batke, Sabrina Szakal, Barnabas Lowther, Jonathan Hao, Fanfan Sarangi, Prabha Branzei, Dana Ulrich, Helle D. Zhao, Xiaolan Concerted and differential actions of two enzymatic domains underlie Rad5 contributions to DNA damage tolerance |
title | Concerted and differential actions of two enzymatic domains underlie Rad5 contributions to DNA damage tolerance |
title_full | Concerted and differential actions of two enzymatic domains underlie Rad5 contributions to DNA damage tolerance |
title_fullStr | Concerted and differential actions of two enzymatic domains underlie Rad5 contributions to DNA damage tolerance |
title_full_unstemmed | Concerted and differential actions of two enzymatic domains underlie Rad5 contributions to DNA damage tolerance |
title_short | Concerted and differential actions of two enzymatic domains underlie Rad5 contributions to DNA damage tolerance |
title_sort | concerted and differential actions of two enzymatic domains underlie rad5 contributions to dna damage tolerance |
topic | Genome Integrity, Repair and Replication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4357696/ https://www.ncbi.nlm.nih.gov/pubmed/25690888 http://dx.doi.org/10.1093/nar/gkv004 |
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