Initiation of a G2/M checkpoint after ultraviolet radiation requires p38 kinase.
Response to genotoxic stress can be considered as a multistage process involving initiation of cell-cycle arrest and maintenance of arrest during DNA repair. Although maintenance of G2/M checkpoints is known to involve Chk1, Chk2/Rad53 and upstream components, the mechanisms involved in its initiation are less well defined. Here we report that p38 kinase has a critical role in the initiation of a G2 delay after ultraviolet radiation. Inhibition of p38 blocks the rapid initiation of this checkpoint in both human and murine cells after ultraviolet radiation. In vitro, p38 binds and phosphorylates Cdc25B at serines 309 and 361, and Cdc25C at serine 216; phosphorylation of these residues is required for binding to 14-3-3 proteins. In vivo, inhibition of p38 prevents both phosphorylation of Cdc25B at serine 309 and 14-3-3 binding after ultraviolet radiation, and mutation of this site is sufficient to inhibit the checkpoint initiation. In contrast, in vivo Cdc25C binding to 14-3-3 is not affected by p38 inhibition after ultraviolet radiation. We propose that regulation of Cdc25B phosphorylation by p38 is a critical event for initiating the G2/M checkpoint after ultraviolet radiation.
Bulavin
D V
DV
Division of Basic Science, National Cancer Intitute, National Institutes of Health, Bethesda, MD 20892, USA.
Higashimoto
Y
Y
Popoff
I J
IJ
Gaarde
W A
WA
Basrur
V
V
Potapova
O
O
Appella
E
E
Fornace
A J
AJ
D048948:14-3-3 Proteins; D000818:Animals; D018797:Cell Cycle Proteins / Q000378:metabolism*; D016195:G2 Phase / Q000235:genetics / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D008938:Mitosis / Q000235:genetics / Q000502:physiology*; D008940:Mitotic Index; D010766:Phosphorylation; D011485:Protein Binding; D012694:Serine / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism; D014466:Ultraviolet Rays; D020687:cdc25 Phosphatases / Q000378:metabolism*; D048051:p38 Mitogen-Activated Protein Kinases
D048948:14-3-3 Proteins; D018797:Cell Cycle Proteins; D012694:Serine; D014446:Tyrosine 3-Monooxygenase; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; C517008:CDC25B protein, human; D020687:cdc25 Phosphatases