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Understanding the role of DNA damage checkpoint kinases in the cellular response to genotoxic stress requires the knowledge of their substrates. Here, we report the use of quantitative phosphoproteomics to identify in vivo kinase substrates of the yeast DNA damage checkpoint kinases Mec1, Tel1, and Rad53 (orthologs of human ATR, ATM, and CHK2, respectively). By analyzing 2,689 phosphorylation sites in wild-type and various kinase-null cells, 62 phosphorylation sites from 55 proteins were found to be controlled by the DNA damage checkpoint. Examination of the dependency of each phosphorylation on Mec1 and Tel1 or Rad53, combined with sequence and biochemical analysis, revealed that many of the identified targets are likely direct substrates of these kinases. In addition to several known targets, 50 previously undescribed targets of the DNA damage checkpoint were identified, suggesting that a wide range of cellular processes is likely regulated by Mec1, Tel1, and Rad53.

Titel:	Proteome-wide identification of in vivo targets of DNA damage checkpoint kinases.
Autor/in / Beteiligte Person:	Smolka, MB ; Albuquerque, CP ; Chen, SH ; Zhou, H
Link:	Full Text Finder (Volltext)
Zeitschrift:	Proceedings of the National Academy of Sciences of the United States of America, Jg. 104 (2007-06-19), Heft 25, S. 10364-9
Veröffentlichung:	Washington, DC : National Academy of Sciences, 2007
Medientyp:	academicJournal
ISSN:	0027-8424 (print)
DOI:	10.1073/pnas.0701622104
Schlagwort:	Cell Cycle Proteins metabolism Checkpoint Kinase 2 Chromatography, Liquid Fungal Proteins metabolism Intracellular Signaling Peptides and Proteins metabolism Methyl Methanesulfonate pharmacology Mutagens pharmacology Protein Kinases metabolism Protein Serine-Threonine Kinases metabolism Saccharomyces cerevisiae genetics Saccharomyces cerevisiae metabolism Saccharomyces cerevisiae Proteins metabolism Substrate Specificity Tandem Mass Spectrometry DNA Damage Fungal Proteins analysis Protein Kinases analysis Proteome analysis
Sonstiges:	Nachgewiesen in: MEDLINE Sprachen: English Publication Type: Journal Article; Research Support, Non-U.S. Gov't Language: English [Proc Natl Acad Sci U S A] 2007 Jun 19; Vol. 104 (25), pp. 10364-9. <i>Date of Electronic Publication: </i>2007 Jun 11. MeSH Terms: DNA Damage* ; Fungal Proteins / analysis ; Protein Kinases / analysis ; Proteome / *analysis ; Cell Cycle Proteins / metabolism ; Checkpoint Kinase 2 ; Chromatography, Liquid ; Fungal Proteins / metabolism ; Intracellular Signaling Peptides and Proteins / metabolism ; Methyl Methanesulfonate / pharmacology ; Mutagens / pharmacology ; Protein Kinases / metabolism ; Protein Serine-Threonine Kinases / metabolism ; Saccharomyces cerevisiae / genetics ; Saccharomyces cerevisiae / metabolism ; Saccharomyces cerevisiae Proteins / metabolism ; Substrate Specificity ; Tandem Mass Spectrometry References: Methods Enzymol. 2002;351:279-96. (PMID: 12073350) ; Proc Natl Acad Sci U S A. 2002 Mar 19;99(6):3746-51. (PMID: 11904430) ; Sci STKE. 2002 Dec 10;2002(162):pe49. (PMID: 12475999) ; Mol Cell Biol. 2003 Feb;23(4):1441-52. (PMID: 12556502) ; Proc Natl Acad Sci U S A. 2003 May 27;100(11):6628-33. (PMID: 12732713) ; Genes Dev. 2003 Jul 15;17(14):1755-67. (PMID: 12865299) ; Nature. 2003 Oct 16;425(6959):737-41. (PMID: 14562106) ; Nature. 2003 Oct 23;425(6960):859-64. (PMID: 14574415) ; DNA Repair (Amst). 2003 Dec 9;2(12):1321-35. (PMID: 14642562) ; EMBO J. 2004 Mar 10;23(5):1188-97. (PMID: 14988729) ; Proc Natl Acad Sci U S A. 2004 Aug 17;101(33):12130-5. (PMID: 15302935) ; Genes Dev. 1990 May;4(5):740-51. (PMID: 2199320) ; Cell. 1993 Dec 17;75(6):1119-27. (PMID: 8261511) ; Cell. 1995 Sep 8;82(5):841-7. (PMID: 7671311) ; Science. 1996 Jan 19;271(5247):357-60. (PMID: 8553072) ; Genes Dev. 1996 Feb 15;10(4):395-406. (PMID: 8600024) ; Proc Natl Acad Sci U S A. 1996 Dec 24;93(26):15075-80. (PMID: 8986766) ; Science. 1997 Dec 19;278(5346):2075-80. (PMID: 9405336) ; Mol Cell. 1998 Aug;2(2):183-9. (PMID: 9734355) ; EMBO J. 1998 Oct 1;17(19):5525-8. (PMID: 9755152) ; EMBO J. 1998 Oct 1;17(19):5679-88. (PMID: 9755168) ; Mol Cell. 2004 Dec 3;16(5):687-700. (PMID: 15574325) ; Nature. 2005 Dec 1;438(7068):679-84. (PMID: 16319894) ; Genes Dev. 2006 Feb 1;20(3):334-44. (PMID: 16452505) ; Cell. 2006 Mar 10;124(5):1069-81. (PMID: 16487579) ; Mol Syst Biol. 2005;1:2005.0017. (PMID: 16729052) ; Cell Cycle. 2006 Jul;5(14):1549-59. (PMID: 16861895) ; Cell. 2006 Nov 3;127(3):635-48. (PMID: 17081983) ; J Cell Biol. 2006 Dec 4;175(5):743-53. (PMID: 17130285) ; DNA Repair (Amst). 2005 Apr 4;4(4):459-68. (PMID: 15725626) ; Mol Cell Proteomics. 2005 Sep;4(9):1358-69. (PMID: 15972895) ; Science. 1999 Nov 5;286(5442):1166-71. (PMID: 10550056) ; J Biol Chem. 1999 Dec 31;274(53):37538-43. (PMID: 10608806) ; Cell. 2000 Jan 7;100(1):113-27. (PMID: 10647936) ; Curr Opin Genet Dev. 2000 Feb;10(1):17-25. (PMID: 10679395) ; Mol Cell Biol. 2000 Jun;20(12):4393-404. (PMID: 10825202) ; Nature. 2000 Dec 21-28;408(6815):1001-4. (PMID: 11140636) ; J Biol Chem. 2001 Aug 3;276(31):29268-74. (PMID: 11387327) ; Nature. 2001 Aug 2;412(6846):557-61. (PMID: 11484058) ; Genes Dev. 2001 Sep 1;15(17):2238-49. (PMID: 11544181) ; Nat Biotechnol. 2002 Mar;20(3):301-5. (PMID: 11875433) ; Nucleic Acids Res. 2006;34(21):6327-36. (PMID: 17108359) ; Mol Cell. 2006 Dec 28;24(6):891-901. (PMID: 17189191) ; J Biol Chem. 2007 Jan 12;282(2):986-95. (PMID: 17114794) ; Annu Rev Genet. 2002;36:617-56. (PMID: 12429704) Grant Information: K22 HG002604 United States HG NHGRI NIH HHS; R01 GM080469 United States GM NIGMS NIH HHS; R01 GM080469-01 United States GM NIGMS NIH HHS Substance Nomenclature: 0 (Cell Cycle Proteins) ; 0 (Fungal Proteins) ; 0 (Intracellular Signaling Peptides and Proteins) ; 0 (Mutagens) ; 0 (Proteome) ; 0 (Saccharomyces cerevisiae Proteins) ; AT5C31J09G (Methyl Methanesulfonate) ; EC 2.7.- (Protein Kinases) ; EC 2.7.1.11 (Checkpoint Kinase 2) ; EC 2.7.11.1 (MEC1 protein, S cerevisiae) ; EC 2.7.11.1 (Protein Serine-Threonine Kinases) ; EC 2.7.11.1 (TEL1 protein, S cerevisiae) ; EC 2.7.12.1 (RAD53 protein, S cerevisiae) Entry Date(s): Date Created: 20070615 Date Completed: 20070815 Latest Revision: 20220331 Update Code: 20240513 PubMed Central ID: PMC1965519

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Proteome-wide identification of in vivo targets of DNA damage checkpoint kinases.