Chk1 is an essential kinase that is regulated by Atr and required for the G(2)/M DNA damage checkpoint.
Chk1, an evolutionarily conserved protein kinase, has been implicated in cell cycle checkpoint control in lower eukaryotes. By gene disruption, we show that CHK1 deficiency results in a severe proliferation defect and death in embryonic stem (ES) cells, and peri-implantation embryonic lethality in mice. Through analysis of a conditional CHK1-deficient cell line, we demonstrate that ES cells lacking Chk1 have a defective G(2)/M DNA damage checkpoint in response to gamma-irradiation (IR). CHK1 heterozygosity modestly enhances the tumorigenesis phenotype of WNT-1 transgenic mice. We show that in human cells, Chk1 is phosphorylated on serine 345 (S345) in response to UV, IR, and hydroxyurea (HU). Overexpression of wild-type Atr enhances, whereas overexpression of the kinase-defective mutant Atr inhibits S345 phosphorylation of Chk1 induced by UV treatment. Taken together, these data indicate that Chk1 plays an essential role in the mammalian DNA damage checkpoint, embryonic development, and tumor suppression, and that Atr regulates Chk1.
Liu
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Q
Howard Hughes Medical Institute, Baylor College of Medicine, Houston, Texas 77030, USA.
Guntuku
S
S
Cui
X S
XS
Matsuoka
S
S
Cortez
D
D
Tamai
K
K
Luo
G
G
Carattini-Rivera
S
S
DeMayo
F
F
Bradley
A
A
Donehower
L A
LA
Elledge
S J
SJ
D000483:Alleles; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis; D064007:Ataxia Telangiectasia Mutated Proteins; D018797:Cell Cycle Proteins; D002478:Cells, Cultured; D000071877:Checkpoint Kinase 1; D004249:DNA Damage; D004260:DNA Repair; D005260:Female; D005434:Flow Cytometry; D016195:G2 Phase; D005720:Gamma Rays; D006579:Heterozygote; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology; D020287:In Situ Nick-End Labeling; D008325:Mammary Neoplasms, Experimental / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D008938:Mitosis / Q000187:drug effects / Q000528:radiation effects; D008957:Models, Genetic; D016296:Mutagenesis; D009368:Neoplasm Transplantation; D015739:Nocodazole / Q000494:pharmacology; D010766:Phosphorylation; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism*; D013234:Stem Cells; D013997:Time Factors; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D014466:Ultraviolet Rays
D000970:Antineoplastic Agents; D018797:Cell Cycle Proteins; D016159:Tumor Suppressor Protein p53; D011494:Protein Kinases; C576897:Atr protein, mouse; C576597:ATR protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; C000605867:CHEK1 protein, human; D000071877:Checkpoint Kinase 1; C000605869:Chek1 protein, mouse; D017346:Protein Serine-Threonine Kinases; D015739:Nocodazole; D006918:Hydroxyurea