Skip to main content

Substrate specificities and identification of putative substrates of ATM kinase family members.

Publication ,  Journal Article
Kim, ST; Lim, DS; Canman, CE; Kastan, MB
Published in: J Biol Chem
December 31, 1999

Ataxia telangiectasia mutated (ATM) phosphorylates p53 protein in response to ionizing radiation, but the complex phenotype of AT cells suggests that it must have other cellular substrates as well. To identify substrates for ATM and the related kinases ATR and DNA-PK, we optimized in vitro kinase assays and developed a rapid peptide screening method to determine general phosphorylation consensus sequences. ATM and ATR require Mn(2+), but not DNA ends or Ku proteins, for optimal in vitro activity while DNA-PKCs requires Mg(2+), DNA ends, and Ku proteins. From p53 peptide mutagenesis analysis, we found that the sequence S/TQ is a minimal essential requirement for all three kinases. In addition, hydrophobic amino acids and negatively charged amino acids immediately NH(2)-terminal to serine or threonine are positive determinants and positively charged amino acids in the region are negative determinants for substrate phosphorylation. We determined a general phosphorylation consensus sequence for ATM and identified putative in vitro targets by using glutathione S-transferase peptides as substrates. Putative ATM in vitro targets include p95/nibrin, Mre11, Brca1, Rad17, PTS, WRN, and ATM (S440) itself. Brca2, phosphatidylinositol 3-kinase, and DNA-5B peptides were phosphorylated specifically by ATR, and DNA Ligase IV is a specific in vitro substrate of DNA-PK.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

December 31, 1999

Volume

274

Issue

53

Start / End Page

37538 / 37543

Location

United States

Related Subject Headings

  • Tumor Suppressor Proteins
  • Substrate Specificity
  • Protein Serine-Threonine Kinases
  • Phosphorylation
  • Phosphatidylinositol 3-Kinases
  • Nuclear Proteins
  • Molecular Sequence Data
  • Humans
  • DNA-Binding Proteins
  • DNA-Activated Protein Kinase
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Kim, S. T., Lim, D. S., Canman, C. E., & Kastan, M. B. (1999). Substrate specificities and identification of putative substrates of ATM kinase family members. J Biol Chem, 274(53), 37538–37543. https://doi.org/10.1074/jbc.274.53.37538
Kim, S. T., D. S. Lim, C. E. Canman, and M. B. Kastan. “Substrate specificities and identification of putative substrates of ATM kinase family members.J Biol Chem 274, no. 53 (December 31, 1999): 37538–43. https://doi.org/10.1074/jbc.274.53.37538.
Kim ST, Lim DS, Canman CE, Kastan MB. Substrate specificities and identification of putative substrates of ATM kinase family members. J Biol Chem. 1999 Dec 31;274(53):37538–43.
Kim, S. T., et al. “Substrate specificities and identification of putative substrates of ATM kinase family members.J Biol Chem, vol. 274, no. 53, Dec. 1999, pp. 37538–43. Pubmed, doi:10.1074/jbc.274.53.37538.
Kim ST, Lim DS, Canman CE, Kastan MB. Substrate specificities and identification of putative substrates of ATM kinase family members. J Biol Chem. 1999 Dec 31;274(53):37538–37543.

Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

December 31, 1999

Volume

274

Issue

53

Start / End Page

37538 / 37543

Location

United States

Related Subject Headings

  • Tumor Suppressor Proteins
  • Substrate Specificity
  • Protein Serine-Threonine Kinases
  • Phosphorylation
  • Phosphatidylinositol 3-Kinases
  • Nuclear Proteins
  • Molecular Sequence Data
  • Humans
  • DNA-Binding Proteins
  • DNA-Activated Protein Kinase