PMID- 10608806
OWN - NLM
STAT- MEDLINE
DCOM- 20000208
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 53
DP  - 1999 Dec 31
TI  - Substrate specificities and identification of putative substrates of ATM kinase
      family members.
PG  - 37538-43
AB  - 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.
FAU - Kim, S T
AU  - Kim ST
AD  - Department of Hematology, St. Jude Children's Research Hospital, Memphis,
      Tennessee 38105, USA.
FAU - Lim, D S
AU  - Lim DS
FAU - Canman, C E
AU  - Canman CE
FAU - Kastan, M B
AU  - Kastan MB
LA  - eng
GR  - CA21765/CA/NCI NIH HHS/United States
GR  - CA71387/CA/NCI NIH HHS/United States
GR  - ES0577/ES/NIEHS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Cell Cycle Proteins)
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Nuclear Proteins)
RN  - 0 (Tumor Suppressor Proteins)
RN  - EC 2.7.1.- (Phosphatidylinositol 3-Kinases)
RN  - EC 2.7.11.1 (ATM protein, human)
RN  - EC 2.7.11.1 (ATR protein, human)
RN  - EC 2.7.11.1 (Ataxia Telangiectasia Mutated Proteins)
RN  - EC 2.7.11.1 (DNA-Activated Protein Kinase)
RN  - EC 2.7.11.1 (PRKDC protein, human)
RN  - EC 2.7.11.1 (Protein-Serine-Threonine Kinases)
SB  - IM
MH  - Amino Acid Sequence
MH  - Ataxia Telangiectasia Mutated Proteins
MH  - Cell Cycle Proteins/metabolism
MH  - Cell Line
MH  - Consensus Sequence
MH  - DNA-Activated Protein Kinase
MH  - *DNA-Binding Proteins
MH  - Humans
MH  - Molecular Sequence Data
MH  - Nuclear Proteins
MH  - Phosphatidylinositol 3-Kinases/metabolism
MH  - Phosphorylation
MH  - Protein-Serine-Threonine Kinases/*metabolism
MH  - Substrate Specificity
MH  - Tumor Suppressor Proteins
EDAT- 1999/12/23 00:00
MHDA- 1999/12/23 00:01
CRDT- 1999/12/23 00:00
PHST- 1999/12/23 00:00 [pubmed]
PHST- 1999/12/23 00:01 [medline]
PHST- 1999/12/23 00:00 [entrez]
AID - 10.1074/jbc.274.53.37538 [doi]
AID - S0021-9258(19)52926-X [pii]
PST - ppublish
SO  - J Biol Chem. 1999 Dec 31;274(53):37538-43. doi: 10.1074/jbc.274.53.37538.