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Mutation of Serine 1333 in the ATR HEAT Repeats Creates a Hyperactive Kinase
Subcellular localization, protein interactions, and post-translational modifications regulate the DNA damage response kinases ATR, ATM, and DNA-PK. During an analysis of putative ATR phosphorylation sites, we found that a single mutation at S1333 creates a hyperactive kinase. In vitro and in cells,...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4047089/ https://www.ncbi.nlm.nih.gov/pubmed/24901225 http://dx.doi.org/10.1371/journal.pone.0099397 |
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author | Luzwick, Jessica W. Nam, Edward A. Zhao, Runxiang Cortez, David |
author_facet | Luzwick, Jessica W. Nam, Edward A. Zhao, Runxiang Cortez, David |
author_sort | Luzwick, Jessica W. |
collection | PubMed |
description | Subcellular localization, protein interactions, and post-translational modifications regulate the DNA damage response kinases ATR, ATM, and DNA-PK. During an analysis of putative ATR phosphorylation sites, we found that a single mutation at S1333 creates a hyperactive kinase. In vitro and in cells, mutation of S1333 to alanine (S1333A-ATR) causes elevated levels of kinase activity with and without the addition of the protein activator TOPBP1. S1333 mutations to glycine, arginine, or lysine also create a hyperactive kinase, while mutation to aspartic acid decreases ATR activity. S1333A-ATR maintains the G(2) checkpoint and promotes completion of DNA replication after transient exposure to replication stress but the less active kinase, S1333D-ATR, has modest defects in both of these functions. While we find no evidence that S1333 is phosphorylated in cultured cells, our data indicate that small changes in the HEAT repeats can have large effects on kinase activity. These mutants may serve as useful tools for future studies of the ATR pathway. |
format | Online Article Text |
id | pubmed-4047089 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-40470892014-06-09 Mutation of Serine 1333 in the ATR HEAT Repeats Creates a Hyperactive Kinase Luzwick, Jessica W. Nam, Edward A. Zhao, Runxiang Cortez, David PLoS One Research Article Subcellular localization, protein interactions, and post-translational modifications regulate the DNA damage response kinases ATR, ATM, and DNA-PK. During an analysis of putative ATR phosphorylation sites, we found that a single mutation at S1333 creates a hyperactive kinase. In vitro and in cells, mutation of S1333 to alanine (S1333A-ATR) causes elevated levels of kinase activity with and without the addition of the protein activator TOPBP1. S1333 mutations to glycine, arginine, or lysine also create a hyperactive kinase, while mutation to aspartic acid decreases ATR activity. S1333A-ATR maintains the G(2) checkpoint and promotes completion of DNA replication after transient exposure to replication stress but the less active kinase, S1333D-ATR, has modest defects in both of these functions. While we find no evidence that S1333 is phosphorylated in cultured cells, our data indicate that small changes in the HEAT repeats can have large effects on kinase activity. These mutants may serve as useful tools for future studies of the ATR pathway. Public Library of Science 2014-06-05 /pmc/articles/PMC4047089/ /pubmed/24901225 http://dx.doi.org/10.1371/journal.pone.0099397 Text en © 2014 Luzwick et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Luzwick, Jessica W. Nam, Edward A. Zhao, Runxiang Cortez, David Mutation of Serine 1333 in the ATR HEAT Repeats Creates a Hyperactive Kinase |
title | Mutation of Serine 1333 in the ATR HEAT Repeats Creates a Hyperactive Kinase |
title_full | Mutation of Serine 1333 in the ATR HEAT Repeats Creates a Hyperactive Kinase |
title_fullStr | Mutation of Serine 1333 in the ATR HEAT Repeats Creates a Hyperactive Kinase |
title_full_unstemmed | Mutation of Serine 1333 in the ATR HEAT Repeats Creates a Hyperactive Kinase |
title_short | Mutation of Serine 1333 in the ATR HEAT Repeats Creates a Hyperactive Kinase |
title_sort | mutation of serine 1333 in the atr heat repeats creates a hyperactive kinase |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4047089/ https://www.ncbi.nlm.nih.gov/pubmed/24901225 http://dx.doi.org/10.1371/journal.pone.0099397 |
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