Defective G2 checkpoint function in cells from individuals with familial cancer syndromes.
The early events in the G2 checkpoint response to ionizing radiation (IR) were analyzed in diploid normal human fibroblasts (NHFs) and fibroblasts from patients with two heritable cancer syndromes. Exposure to gamma-radiation of asynchronously growing NHFs resulted in a rapid reduction in the number of cells in mitosis (G2 delay) and was accompanied by a quantitatively similar reduction in the p34CDC2/cyclin B in vitro histone H1 kinase activity as compared with sham-treated controls. This G2 delay was strong by 1 h following exposure to IR, maximal by 2 h, and was accompanied by an accumulation of tyrosine-phosphorylated p34CDC2 molecules. In contrast, fibroblasts from individuals with ataxia telangiectasia displayed significantly less reduction of the mitotic index or histone H1 kinase activity after IR. Low passage fibroblasts from individuals with Li-Fraumeni syndrome having one wild-type and one mutated p53 allele were similar to NHFs in their immediate G2 checkpoint response to IR, as were NHFs expressing the human papilloma virus type 16 E6 gene product (functionally inactivating p53) and low passage cells from p53-deficient mouse embryos. However, the p53-deficient fibroblasts were genomically unstable and became defective in their early G2 checkpoint response to IR. Furthermore, immortal Li-Fraumeni syndrome fibroblasts lacking wild-type p53 displayed an attenuated G2 checkpoint response. These results link the early events in G2 checkpoint response to IR in NHFs with a rapid inhibition of p34CDC2/cyclin B protein kinase activity and demonstrate that while not required for this immediate G2 delay, lack of p53 can lead to subsequent genetic alterations that result in defective G2 checkpoint function.
Paules
R S
RS
Growth Control and Cancer Group, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709, USA.
Levedakou
E N
EN
Wilson
S J
SJ
Innes
C L
CL
Rhodes
N
N
Tlsty
T D
TD
Galloway
D A
DA
Donehower
L A
LA
Tainsky
M A
MA
Kaufmann
W K
WK
D001260:Ataxia Telangiectasia / Q000235:genetics* / Q000473:pathology; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology; D016368:Frameshift Mutation; D016195:G2 Phase / Q000235:genetics* / Q000528:radiation effects; D005720:Gamma Rays; D016158:Genes, p53; D006801:Humans; D007231:Infant, Newborn; D007700:Kinetics; D016864:Li-Fraumeni Syndrome / Q000235:genetics* / Q000473:pathology; D008938:Mitosis; D008940:Mitotic Index / Q000528:radiation effects; D006656:Protamine Kinase / Q000378:metabolism; D012016:Reference Values; D013997:Time Factors
D006656:Protamine Kinase