Concerted and differential actions of two enzymatic domains underlie Rad5 contributions to DNA damage tolerance Journal Article
| Authors: | Choi, K.; Batke, S.; Szakal, B.; Lowther, J.; Hao, F.; Sarangi, P.; Branzei, D.; Ulrich, H. D.; Zhao, X. |
| Article Title: | Concerted and differential actions of two enzymatic domains underlie Rad5 contributions to DNA damage tolerance |
| Abstract: | 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 polyubiquitination 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 helicasemediated 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. © The Author(s) 2015. |
| Keywords: | survival; controlled study; unclassified drug; nonhuman; genetic analysis; protein domain; protein motif; dna recombination; dna damage; ubiquitin protein ligase; protein interaction; in vitro study; enzyme activity; ubiquitination; plasmid; stress; saccharomycetales; adenosine triphosphate; cycline; adenosine triphosphatase; dna helicase; fungal protein; fungal strain; protein rad5; priority journal; article |
| Journal Title: | Nucleic Acids Research |
| Volume: | 43 |
| Issue: | 5 |
| ISSN: | 0305-1048 |
| Publisher: | Oxford University Press |
| Date Published: | 2015-03-11 |
| Start Page: | 2666 |
| End Page: | 2677 |
| Language: | English |
| DOI: | 10.1093/nar/gkv004 |
| PROVIDER: | scopus |
| PMCID: | PMC4357696 |
| PUBMED: | 25690888 |
| DOI/URL: | |
| Notes: | Export Date: 2 September 2015 -- Source: Scopus |
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