Purification and characterization of ATM from human placenta

Chan, D., Son, S-C., Block, W., Ye, R., Khanna, K. K., Wold, M. S., Douglas, P. J. G., Goodarzi, A. A., Pelley, J., Taya, Y., Lavin, M. F. and Lees-Miller, S. P. (2000) Purification and characterization of ATM from human placenta. Journal of Biological Chemistry, 275 11: 7803-7810. doi:10.1074/jbc.275.11.7803

Attached Files (Some files may be inaccessible until you login with your UQ eSpace credentials)
Name Description MIMEType Size Downloads
UQ140418_OA.pdf Full text (open access) application/pdf 303.88KB 0

Author Chan, D.
Son, S-C.
Block, W.
Ye, R.
Khanna, K. K.
Wold, M. S.
Douglas, P. J. G.
Goodarzi, A. A.
Pelley, J.
Taya, Y.
Lavin, M. F.
Lees-Miller, S. P.
Title Purification and characterization of ATM from human placenta
Journal name Journal of Biological Chemistry   Check publisher's open access policy
ISSN 0021-9258
Publication date 2000-03-16
Sub-type Article (original research)
DOI 10.1074/jbc.275.11.7803
Open Access Status File (Publisher version)
Volume 275
Issue 11
Start page 7803
End page 7810
Total pages 8
Place of publication Bethesda, U.S.A.
Publisher American Society for Biochemistry & Molecular Biology
Language eng
Subject C1
321015 Oncology and Carcinogenesis
730108 Cancer and related disorders
Abstract ATM is mutated in the human genetic disorder ataxia telangiectasia, which is characterized by ataxia, immune defects, and cancer predisposition. Cells that lack ATM exhibit delayed up-regulation of p53 in response to ionizing radiation. Serine 15 of p53 is phosphorylated in vivo in response to ionizing radiation, and antibodies to ATM immunoprecipitate a protein kinase activity that, in the presence of manganese, phosphorylates p53 at serine 15. Immunoprecipitates of ATM also phosphorylate PHAS-I in a manganese-dependent manner. Here we have purified ATM from human cells using nine chromatographic steps. Highly purified ATM phosphorylated PHAS-I, the 32-kDa subunit of RPA, serine 15 of p53, and Chk2 in vitro. The majority of the ATM phosphorylation sites in Chk2 were located in the amino-terminal 57 amino acids. In each case, phosphorylation was strictly dependent on manganese. ATM protein kinase activity was inhibited by wortmannin with an IC50 of approximately 100 nM. Phosphorylation of RPA, but not p53, Chk2, or PHAS-I, was stimulated by DNA. The related protein, DNA-dependent protein kinase catalytic subunit, also phosphorylated PHAS-I, RPA, and Chk2 in the presence of manganese, suggesting that the requirement for manganese is a characteristic of this class of enzyme.
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Medicine Publications
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 93 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 94 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Tue, 10 Jun 2008, 21:29:06 EST