ATP activates ataxia-telangiectasia mutated (ATM) in vitro - Importance of autophosphorylation

Kozlov, Sergei, Gueven, Nuri, Keating, Katherine, Ramsay, Jonathan and Lavin, Martin F. (2003) ATP activates ataxia-telangiectasia mutated (ATM) in vitro - Importance of autophosphorylation. Journal of Biological Chemistry, 278 11: 9309-9317. doi:10.1074/jbc.M300003200

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Author Kozlov, Sergei
Gueven, Nuri
Keating, Katherine
Ramsay, Jonathan
Lavin, Martin F.
Title ATP activates ataxia-telangiectasia mutated (ATM) in vitro - Importance of autophosphorylation
Journal name Journal of Biological Chemistry   Check publisher's open access policy
ISSN 0021-9258
Publication date 2003-03-14
Sub-type Article (original research)
DOI 10.1074/jbc.M300003200
Open Access Status File (Publisher version)
Volume 278
Issue 11
Start page 9309
End page 9317
Total pages 9
Editor H. Tabor
Place of publication Bethesda, U.S.A.
Publisher American Society for Biochemistry & Molecular Biology
Language eng
Subject C1
320305 Medical Biochemistry - Proteins and Peptides
730107 Inherited diseases (incl. gene therapy)
0304 Medicinal and Biomolecular Chemistry
Abstract Ataxia-telangiectasia Mutated (ATM), mutated in the human disorder ataxia-telangiectasia, is rapidly activated by DNA double strand breaks. The mechanism of activation remains unresolved, and it is uncertain whether autophosphorylation contributes to activation. We describe an in vitro immunoprecipitation system demonstrating activation of ATM kinase from unirradiated extracts by preincubation with ATP. Activation is both time- and ATP concentration-dependent, other nucleotides fail to activate ATM, and DNA is not required. ATP activation is specific for ATM since it is not observed with kinase-dead ATM, it requires Mn2+, and it is inhibited by wortmannin. Exposure of activated ATM to phosphatase abrogates activity, and repeat cycles of ATP and phosphatase treatment reveal a requirement for autophosphorylation in the activation process. Phosphopeptide mapping revealed similarities between the patterns of autophosphorylation for irradiated and ATP-treated ATM. Caffeine inhibited ATM kinase activity for substrates but did not interfere with ATM autophosphorylation. ATP failed to activate either A-T and rad3-related protein (ATR) or DNA-dependent protein kinase under these conditions, supporting the specificity for ATM. These data demonstrate that ATP can specifically induce activation of ATM by a mechanism involving autophosphorylation. The relationship of this activation to DNA damage activation remains unclear but represents a useful model for understanding in vivo activation.
Keyword Biochemistry & Molecular Biology
Dependent Protein-kinase
Double-strand Breaks
S-phase Checkpoint
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
2004 Higher Education Research Data Collection
School of Medicine Publications
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Citation counts: TR Web of Science Citation Count  Cited 61 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 68 times in Scopus Article | Citations
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Created: Wed, 15 Aug 2007, 11:25:30 EST