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Ubc4 and Not4 Regulate Steady-State Levels of DNA Polymerase-α to Promote Efficient and Accurate DNA Replication

The accurate duplication of chromosomal DNA is required to maintain genomic integrity. However, from an evolutionary point of view, a low mutation rate during DNA replication is desirable. One way to strike the right balance between accuracy and limited mutagenesis is to use a DNA polymerase that la...

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Autores principales: Haworth, Justin, Alver, Robert C., Anderson, Melissa, Bielinsky, Anja-Katrin
Formato: Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2938386/
https://www.ncbi.nlm.nih.gov/pubmed/20660159
http://dx.doi.org/10.1091/mbc.E09-06-0452
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author Haworth, Justin
Alver, Robert C.
Anderson, Melissa
Bielinsky, Anja-Katrin
author_facet Haworth, Justin
Alver, Robert C.
Anderson, Melissa
Bielinsky, Anja-Katrin
author_sort Haworth, Justin
collection PubMed
description The accurate duplication of chromosomal DNA is required to maintain genomic integrity. However, from an evolutionary point of view, a low mutation rate during DNA replication is desirable. One way to strike the right balance between accuracy and limited mutagenesis is to use a DNA polymerase that lacks proofreading activity but contributes to DNA replication in a very restricted manner. DNA polymerase-α fits this purpose exactly, but little is known about its regulation at the replication fork. Minichromosome maintenance protein (Mcm) 10 regulates the stability of the catalytic subunit of pol-α in budding yeast and human cells. Cdc17, the catalytic subunit of pol-α in yeast, is rapidly degraded after depletion of Mcm10. Here we show that Ubc4 and Not4 are required for Cdc17 destabilization. Disruption of Cdc17 turnover resulted in sensitivity to hydroxyurea, suggesting that this pathway is important for DNA replication. Furthermore, overexpression of Cdc17 in ubc4 and not4 mutants caused slow growth and synthetic dosage lethality, respectively. Our data suggest that Cdc17 levels are very tightly regulated through the opposing forces of Ubc4 and Not4 (destabilization) and Mcm10 (stabilization). We conclude that regular turnover of Cdc17 via Ubc4 and Not4 is required for proper cell proliferation.
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spelling pubmed-29383862010-11-30 Ubc4 and Not4 Regulate Steady-State Levels of DNA Polymerase-α to Promote Efficient and Accurate DNA Replication Haworth, Justin Alver, Robert C. Anderson, Melissa Bielinsky, Anja-Katrin Mol Biol Cell Articles The accurate duplication of chromosomal DNA is required to maintain genomic integrity. However, from an evolutionary point of view, a low mutation rate during DNA replication is desirable. One way to strike the right balance between accuracy and limited mutagenesis is to use a DNA polymerase that lacks proofreading activity but contributes to DNA replication in a very restricted manner. DNA polymerase-α fits this purpose exactly, but little is known about its regulation at the replication fork. Minichromosome maintenance protein (Mcm) 10 regulates the stability of the catalytic subunit of pol-α in budding yeast and human cells. Cdc17, the catalytic subunit of pol-α in yeast, is rapidly degraded after depletion of Mcm10. Here we show that Ubc4 and Not4 are required for Cdc17 destabilization. Disruption of Cdc17 turnover resulted in sensitivity to hydroxyurea, suggesting that this pathway is important for DNA replication. Furthermore, overexpression of Cdc17 in ubc4 and not4 mutants caused slow growth and synthetic dosage lethality, respectively. Our data suggest that Cdc17 levels are very tightly regulated through the opposing forces of Ubc4 and Not4 (destabilization) and Mcm10 (stabilization). We conclude that regular turnover of Cdc17 via Ubc4 and Not4 is required for proper cell proliferation. The American Society for Cell Biology 2010-09-15 /pmc/articles/PMC2938386/ /pubmed/20660159 http://dx.doi.org/10.1091/mbc.E09-06-0452 Text en © 2010 by The American Society for Cell Biology This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).
spellingShingle Articles
Haworth, Justin
Alver, Robert C.
Anderson, Melissa
Bielinsky, Anja-Katrin
Ubc4 and Not4 Regulate Steady-State Levels of DNA Polymerase-α to Promote Efficient and Accurate DNA Replication
title Ubc4 and Not4 Regulate Steady-State Levels of DNA Polymerase-α to Promote Efficient and Accurate DNA Replication
title_full Ubc4 and Not4 Regulate Steady-State Levels of DNA Polymerase-α to Promote Efficient and Accurate DNA Replication
title_fullStr Ubc4 and Not4 Regulate Steady-State Levels of DNA Polymerase-α to Promote Efficient and Accurate DNA Replication
title_full_unstemmed Ubc4 and Not4 Regulate Steady-State Levels of DNA Polymerase-α to Promote Efficient and Accurate DNA Replication
title_short Ubc4 and Not4 Regulate Steady-State Levels of DNA Polymerase-α to Promote Efficient and Accurate DNA Replication
title_sort ubc4 and not4 regulate steady-state levels of dna polymerase-α to promote efficient and accurate dna replication
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2938386/
https://www.ncbi.nlm.nih.gov/pubmed/20660159
http://dx.doi.org/10.1091/mbc.E09-06-0452
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