Requirement of ATM-dependent phosphorylation of brca1 in the DNA damage response to double-strand breaks.

The Brca1 (breast cancer gene 1) tumor suppressor protein is phosphorylated in response to DNA damage. Results from this study indicate that the checkpoint protein kinase ATM (mutated in ataxia telangiectasia) was required for phosphorylation of Brca1 in response to ionizing radiation. ATM resides in a complex with Brca1 and phosphorylated Brca1 in vivo and in vitro in a region that contains clusters of serine-glutamine residues. Phosphorylation of this domain appears to be functionally important because a mutated Brca1 protein lacking two phosphorylation sites failed to rescue the radiation hypersensitivity of a Brca1-deficient cell line. Thus, phosphorylation of Brca1 by the checkpoint kinase ATM may be critical for proper responses to DNA double-strand breaks and may provide a molecular explanation for the role of ATM in breast cancer.

[1]  A. Ashworth,et al.  Tumorigenesis and a DNA repair defect in mice with a truncating Brca2 mutation , 1997, Nature Genetics.

[2]  H. Gardner,et al.  Developmental expression of Brca2 colocalizes with Brca1 and is associated with proliferation and differentiation in multiple tissues. , 1997, Developmental biology.

[3]  Heinz Ruffner,et al.  BRCA1 Is Phosphorylated at Serine 1497 In Vivo at a Cyclin-Dependent Kinase 2 Phosphorylation Site , 1999, Molecular and Cellular Biology.

[4]  J. Minna,et al.  Characterization of a breast cancer cell line derived from a germ-line BRCA1 mutation carrier. , 1998, Cancer research.

[5]  T. Ouchi,et al.  The BRCA2 gene product functionally interacts with p53 and RAD51. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[6]  F. Couch,et al.  Stable interaction between the products of the BRCA1 and BRCA2 tumor suppressor genes in mitotic and meiotic cells. , 1998, Molecular cell.

[7]  A. Hinnebusch,et al.  Identification of phosphorylation sites in proteins separated by polyacrylamide gel electrophoresis. , 1998, Analytical chemistry.

[8]  P. Leder,et al.  Pleiotropic defects in ataxia-telangiectasia protein-deficient mice. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[9]  Stephen J. Elledge,et al.  The univector plasmid-fusion system, a method for rapid construction of recombinant DNA without restriction enzymes , 1998, Current Biology.

[10]  Stephen J. Elledge,et al.  Cell Cycle Checkpoints: Preventing an Identity Crisis , 1996, Science.

[11]  A. Berchuck,et al.  Cell cycle control of BRCA2. , 1996, Cancer research.

[12]  T. Pandita,et al.  The contribution of DNA and chromosome repair deficiencies to the radiosensitivity of ataxia-telangiectasia. , 1992, Radiation research.

[13]  C. Deng,et al.  A targeted disruption of the murine Brca1 gene causes γ-irradiation hypersensitivity and genetic instability , 1998, Oncogene.

[14]  Bert Vogelstein,et al.  Gatekeepers and caretakers , 1997, Nature.

[15]  M. Swift,et al.  Molecular genotyping shows that ataxia-telangiectasia heterozygotes are predisposed to breast cancer. , 1996, Cancer genetics and cytogenetics.

[16]  B. Ponder,et al.  Involvement of Brca2 in DNA repair. , 1998, Molecular cell.

[17]  Yosef Shiloh,et al.  Recombinant ATM protein complements the cellular A-T phenotype , 1997, Oncogene.

[18]  F. Alt,et al.  RAB22 and RAB163/mouse BRCA2: proteins that specifically interact with the RAD51 protein. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[19]  A. Aurias,et al.  Breast cancer risk in ataxia telangiectasia (AT) heterozygotes: haplotype study in French AT families , 1999, British Journal of Cancer.

[20]  Ralph Scully,et al.  Dynamic Changes of BRCA1 Subnuclear Location and Phosphorylation State Are Initiated by DNA Damage , 1997, Cell.

[21]  A. Berchuck,et al.  BRCA1 expression is induced before DNA synthesis in both normal and tumor-derived breast cells. , 1996, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[22]  Y Taya,et al.  DNA damage induces phosphorylation of the amino terminus of p53. , 1997, Genes & development.

[23]  H. Hanafusa,et al.  Evidence for a transcriptional activation function of BRCA1 C-terminal region. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Y. Shiloh,et al.  ATM: from gene to function. , 1998, Human molecular genetics.

[25]  X. Wang,et al.  Centrosome amplification and a defective G2-M cell cycle checkpoint induce genetic instability in BRCA1 exon 11 isoform-deficient cells. , 1999, Molecular cell.

[26]  J. Rommens,et al.  The complete BRCA2 gene and mutations in chromosome 13q-linked kindreds , 1996, Nature Genetics.

[27]  A. Blasina,et al.  A human homologue of the checkpoint kinase Cds1 directly inhibits Cdc25 phosphatase , 1999, Current Biology.

[28]  H. Piwnica-Worms,et al.  A human Cds1-related kinase that functions downstream of ATM protein in the cellular response to DNA damage. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[29]  D. Baltimore,et al.  Targeted disruption of ATM leads to growth retardation, chromosomal fragmentation during meiosis, immune defects, and thymic lymphoma. , 1996, Genes & development.

[30]  Y. Chen,et al.  BRCA1 is a 220-kDa nuclear phosphoprotein that is expressed and phosphorylated in a cell cycle-dependent manner. , 1996, Cancer research.

[31]  Y. Shiloh,et al.  Interaction between ATM protein and c-Abl in response to DNA damage , 1997, Nature.

[32]  D. Bentley,et al.  Identification of the breast cancer susceptibility gene BRCA2 , 1995, Nature.

[33]  C. Badie,et al.  Hypersensitivity of ataxia telangiectasia fibroblasts to ionizing radiation is associated with a repair deficiency of DNA double-strand breaks. , 1997, International journal of radiation biology.

[34]  M. Hannan,et al.  Fibroblasts from ataxia telangiectasia (AT) and AT heterozygotes show an enhanced level of residual DNA double-strand breaks after low dose-rate gamma-irradiation as assayed by pulsed field gel electrophoresis. , 1991, International journal of radiation biology.

[35]  S. Carr,et al.  Mammalian Chk2 is a downstream effector of the ATM-dependent DNA damage checkpoint pathway , 1999, Oncogene.

[36]  C. Moskaluk,et al.  Germline BRCA2 gene mutations in patients with apparently sporadic pancreatic carcinomas. , 1996, Cancer research.

[37]  P. Bartel,et al.  RAD51 Interacts with the Evolutionarily Conserved BRC Motifs in the Human Breast Cancer Susceptibility Gene brca2 * , 1997, The Journal of Biological Chemistry.

[38]  Yonghong Xiao,et al.  Association of BRCA1 with Rad51 in Mitotic and Meiotic Cells , 1997, Cell.

[39]  T. Mak,et al.  Brca2 is required for embryonic cellular proliferation in the mouse. , 1997, Genes & development.

[40]  H. Ruffner,et al.  BRCA1 is a cell cycle-regulated nuclear phosphoprotein. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[41]  T. Ludwig,et al.  Targeted mutations of breast cancer susceptibility gene homologs in mice: lethal phenotypes of Brca1, Brca2, Brca1/Brca2, Brca1/p53, and Brca2/p53 nullizygous embryos. , 1997, Genes & development.

[42]  K. Biemann.  Contributions of mass spectrometry to peptide and protein structure. , 1988, Biomedical & environmental mass spectrometry.

[43]  S. Elledge,et al.  Linkage of ATM to cell cycle regulation by the Chk2 protein kinase. , 1998, Science.

[44]  Y Taya,et al.  Activation of the ATM kinase by ionizing radiation and phosphorylation of p53. , 1998, Science.

[45]  M. Lovett,et al.  A single ataxia telangiectasia gene with a product similar to PI-3 kinase. , 1995, Science.

[46]  F. Collins,et al.  Human BRCA1 inhibits growth in yeast: potential use in diagnostic testing. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[47]  G. Eichele,et al.  Embryonic lethality and radiation hypersensitivity mediated by Rad51 in mice lacking Brca2 , 1997, Nature.

[48]  A. Schäffer,et al.  Atm haploinsufficiency results in increased sensitivity to sublethal doses of ionizing radiation in mice , 1999, Nature Genetics.

[49]  R. Jensen,et al.  BRCA1 Expression Restores Radiation Resistance in BRCA1-defective Cancer Cells through Enhancement of Transcription-coupled DNA Repair* , 1999, The Journal of Biological Chemistry.

[50]  Y. Shiloh,et al.  Targeting double-strand breaks to replicating DNA identifies a subpathway of DSB repair that is defective in ataxia-telangiectasia cells. , 1999, Biochemical and biophysical research communications.

[51]  D. Baltimore,et al.  Ataxia telangiectasia mutant protein activates c-Abl tyrosine kinase in response to ionizing radiation , 1997, Nature.

[52]  L. Bennett,et al.  Brca1 and Brca2 expression patterns in mitotic and meiotic cells of mice , 1998, Oncogene.

[53]  Y. Houvras,et al.  Arrest of the cell cycle by the tumour-suppressor BRCA1 requires the CDK-inhibitor p21WAF1/CiPl , 1997, Nature.