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Requirement of the FATC domain of protein kinase Tel1 for localization to DNA ends and target protein recognition
Two large phosphatidylinositol 3-kinase–related protein kinases (PIKKs), ATM and ATR, play a central role in the DNA damage response pathway. PIKKs contain a highly conserved extreme C-terminus called the FRAP-ATM-TRRAP-C-terminal (FATC) domain. In budding yeast, ATM and ATR correspond to Tel1 and M...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4591692/ https://www.ncbi.nlm.nih.gov/pubmed/26246601 http://dx.doi.org/10.1091/mbc.E15-05-0259 |
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author | Ogi, Hiroo Goto, Greicy H. Ghosh, Avik Zencir, Sevil Henry, Everett Sugimoto, Katsunori |
author_facet | Ogi, Hiroo Goto, Greicy H. Ghosh, Avik Zencir, Sevil Henry, Everett Sugimoto, Katsunori |
author_sort | Ogi, Hiroo |
collection | PubMed |
description | Two large phosphatidylinositol 3-kinase–related protein kinases (PIKKs), ATM and ATR, play a central role in the DNA damage response pathway. PIKKs contain a highly conserved extreme C-terminus called the FRAP-ATM-TRRAP-C-terminal (FATC) domain. In budding yeast, ATM and ATR correspond to Tel1 and Mec1, respectively. In this study, we characterized functions of the FATC domain of Tel1 by introducing substitution or truncation mutations. One substitution mutation, termed tel1-21, and a truncation mutation, called tel1-ΔC, did not significantly affect the expression level. The tel1-21 mutation impaired the cellular response to DNA damage and conferred moderate telomere maintenance defect. In contrast, the tel1-ΔC mutation behaved like a null mutation, conferring defects in both DNA damage response and telomere maintenance. Tel1-21 protein localized to DNA ends as effectively as wild-type Tel1 protein, whereas Tel1-ΔC protein failed. Introduction of a hyperactive TEL1-hy mutation suppressed the tel1-21 mutation but not the tel1-ΔC mutation. In vitro analyses revealed that both Tel1-21 and Tel1-ΔC proteins undergo efficient autophosphorylation but exhibit decreased kinase activities toward the exogenous substrate protein, Rad53. Our results show that the FATC domain of Tel1 mediates localization to DNA ends and contributes to phosphorylation of target proteins. |
format | Online Article Text |
id | pubmed-4591692 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-45916922015-12-16 Requirement of the FATC domain of protein kinase Tel1 for localization to DNA ends and target protein recognition Ogi, Hiroo Goto, Greicy H. Ghosh, Avik Zencir, Sevil Henry, Everett Sugimoto, Katsunori Mol Biol Cell Articles Two large phosphatidylinositol 3-kinase–related protein kinases (PIKKs), ATM and ATR, play a central role in the DNA damage response pathway. PIKKs contain a highly conserved extreme C-terminus called the FRAP-ATM-TRRAP-C-terminal (FATC) domain. In budding yeast, ATM and ATR correspond to Tel1 and Mec1, respectively. In this study, we characterized functions of the FATC domain of Tel1 by introducing substitution or truncation mutations. One substitution mutation, termed tel1-21, and a truncation mutation, called tel1-ΔC, did not significantly affect the expression level. The tel1-21 mutation impaired the cellular response to DNA damage and conferred moderate telomere maintenance defect. In contrast, the tel1-ΔC mutation behaved like a null mutation, conferring defects in both DNA damage response and telomere maintenance. Tel1-21 protein localized to DNA ends as effectively as wild-type Tel1 protein, whereas Tel1-ΔC protein failed. Introduction of a hyperactive TEL1-hy mutation suppressed the tel1-21 mutation but not the tel1-ΔC mutation. In vitro analyses revealed that both Tel1-21 and Tel1-ΔC proteins undergo efficient autophosphorylation but exhibit decreased kinase activities toward the exogenous substrate protein, Rad53. Our results show that the FATC domain of Tel1 mediates localization to DNA ends and contributes to phosphorylation of target proteins. The American Society for Cell Biology 2015-10-01 /pmc/articles/PMC4591692/ /pubmed/26246601 http://dx.doi.org/10.1091/mbc.E15-05-0259 Text en © 2015 Ogi, Goto, Ghosh, et al. 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). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. |
spellingShingle | Articles Ogi, Hiroo Goto, Greicy H. Ghosh, Avik Zencir, Sevil Henry, Everett Sugimoto, Katsunori Requirement of the FATC domain of protein kinase Tel1 for localization to DNA ends and target protein recognition |
title | Requirement of the FATC domain of protein kinase Tel1 for localization to DNA ends and target protein recognition |
title_full | Requirement of the FATC domain of protein kinase Tel1 for localization to DNA ends and target protein recognition |
title_fullStr | Requirement of the FATC domain of protein kinase Tel1 for localization to DNA ends and target protein recognition |
title_full_unstemmed | Requirement of the FATC domain of protein kinase Tel1 for localization to DNA ends and target protein recognition |
title_short | Requirement of the FATC domain of protein kinase Tel1 for localization to DNA ends and target protein recognition |
title_sort | requirement of the fatc domain of protein kinase tel1 for localization to dna ends and target protein recognition |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4591692/ https://www.ncbi.nlm.nih.gov/pubmed/26246601 http://dx.doi.org/10.1091/mbc.E15-05-0259 |
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