NFBD1/MDC1 associates with p53 and regulates its function at the crossroad between cell survival and death in response to DNA damage.
NFBD1/MDC1, which belongs to the BRCT superfamily, has an anti-apoptotic activity and contributes to the early cellular responses to DNA damage. Here we found that NFBD1 protects cells from apoptotic cell death by inhibiting phosphorylation of p53 at Ser-15 under steady state as well as early phase of DNA damage, thereby blocking its transcriptional and pro-apoptotic activities. During late phase of DNA damage, a remarkable reduction of NFBD1 was observed in dying but not in surviving A549 cells bearing wild-type p53. Small interference RNA-mediated knockdown of the endogenous NFBD1 resulted in an increase in sensitivity to adriamycin in A549 cells but not in p53-deficient H1299 cells. Immunoprecipitation and luciferase reporter analyses demonstrated that NFBD1 binds to the NH(2)-terminal region of p53 and strongly inhibits its transcriptional activity. Additionally, BRCT domains, which can interact with p53, reduced the adriamycin-induced phosphorylation levels of p53 at Ser-15 and also suppressed the transcriptional activity of p53. Thus, our present findings strongly suggest that NFBD1 plays an important role in the decision of cell survival and death after DNA damage through the regulation of p53.
Nakanishi
Mitsuru
M
Division of Biochemistry, Chiba Cancer Center Research Institute, Chiba 260-8717, Japan.
Ozaki
Toshinori
T
Yamamoto
Hideki
H
Hanamoto
Takayuki
T
Kikuchi
Hironobu
H
Furuya
Kazushige
K
Asaka
Masahiro
M
Delia
Domenico
D
Nakagawara
Akira
A
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D017209:Apoptosis; D019556:COS Cells; D018797:Cell Cycle Proteins; D016923:Cell Death; D045744:Cell Line, Tumor; D002470:Cell Survival; D002522:Chlorocebus aethiops; D004249:DNA Damage; D004268:DNA-Binding Proteins / Q000502:physiology; D015972:Gene Expression Regulation, Neoplastic; D016158:Genes, p53; D006801:Humans; D008954:Models, Biological; D009687:Nuclear Proteins / Q000502:physiology*; D034622:RNA Interference; D015534:Trans-Activators / Q000502:physiology*; D015533:Transcriptional Activation
D048868:Adaptor Proteins, Signal Transducing; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; C413163:MDC1 protein, human; D009687:Nuclear Proteins; D015534:Trans-Activators