Functional interaction of p53 and BLM DNA helicase in apoptosis.
The Bloom syndrome (BS) protein, BLM, is a member of the RecQ DNA helicase family that also includes the Werner syndrome protein, WRN. Inherited mutations in these proteins are associated with cancer predisposition of these patients. We recently discovered that cells from Werner syndrome patients displayed a deficiency in p53-mediated apoptosis and WRN binds to p53. Here, we report that analogous to WRN, BLM also binds to p53 in vivo and in vitro, and the C-terminal domain of p53 is responsible for the interaction. p53-mediated apoptosis is defective in BS fibroblasts and can be rescued by expression of the normal BLM gene. Moreover, lymphoblastoid cell lines (LCLs) derived from BS donors are resistant to both gamma-radiation and doxorubicin-induced cell killing, and sensitivity can be restored by the stable expression of normal BLM. In contrast, BS cells have a normal Fas-mediated apoptosis, and in response to DNA damage normal accumulation of p53, normal induction of p53 responsive genes, and normal G(1)-S and G(2)-M cell cycle arrest. BLM localizes to nuclear foci referred to as PML nuclear bodies (NBs). Cells from Li-Fraumeni syndrome patients carrying p53 germline mutations and LCLs lacking a functional p53 have a decreased accumulation of BLM in NBs, whereas isogenic lines with functional p53 exhibit normal accumulation. Certain BLM mutants (C1055S or Delta133-237) that have a reduced ability to localize to the NBs when expressed in normal cells can impair the localization of wild type BLM to NBs and block p53-mediated apoptosis, suggesting a dominant-negative effect. Taken together, our results indicate both a novel mechanism of p53 function by which p53 mediates nuclear trafficking of BLM to NBs and the cooperation of p53 and BLM to induce apoptosis.
Wang
X W
XW
Laboratory of Human Carcinogenesis, Center for Cancer Research, NCI, National Institutes of Health, Bethesda, Maryland 20892, USA.
Tseng
A
A
Ellis
N A
NA
Spillare
E A
EA
Linke
S P
SP
Robles
A I
AI
Seker
H
H
Yang
Q
Q
Hu
P
P
Beresten
S
S
Bemmels
N A
NA
Garfield
S
S
Harris
C C
CC
D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000378:metabolism*; D017209:Apoptosis / Q000502:physiology* / Q000528:radiation effects; D001665:Binding Sites; D001816:Bloom Syndrome / Q000201:enzymology* / Q000235:genetics; D002453:Cell Cycle / Q000502:physiology*; D002460:Cell Line; D002467:Cell Nucleus / Q000502:physiology; D002470:Cell Survival; D004249:DNA Damage; D004265:DNA Helicases / Q000737:chemistry* / Q000378:metabolism*; D004307:Dose-Response Relationship, Radiation; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology / Q000528:radiation effects; D019084:Fluorescent Antibody Technique, Indirect; D005720:Gamma Rays; D017930:Genes, Reporter; D006801:Humans; D053484:RecQ Helicases; D011994:Recombinant Proteins / Q000378:metabolism; D012016:Reference Values; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000378:metabolism*
D011994:Recombinant Proteins; D016159:Tumor Suppressor Protein p53; D000251:Adenosine Triphosphatases; C097151:Bloom syndrome protein; D004265:DNA Helicases; D053484:RecQ Helicases