Redox regulation of Cdc25C.
The Cdc25 family of dual specific phosphatases are critical components of cell cycle progression and checkpoint control. Certain stresses such as ultraviolet light stimulate the rapid and selective destruction of Cdc25A protein through a Chk1 protein kinase-dependent pathway. We demonstrate that in contrast to cellular stresses previously examined, hydrogen peroxide exposure affects Cdc25C but not Cdc25A levels. Pharmacological inhibition of Chk1 activity or a mutant of Cdc25C that lacks the Chk1 phosphorylation site still undergoes degradation in response to oxidants. We also demonstrate that in vitro hydrogen peroxide stimulates an intramolecular disulfide bond between the active site cysteine at position 377 and another invariant cysteine at position 330. The in vivo stability of Cdc25C is substantially reduced by the mutation of either of these two cysteine residues. In contrast, a double (C2) mutant of both cysteine 330 and cysteine 377 results in a protein that is more stable than wild type Cdc25C and is resistant to oxidative stress-induced degradation. In addition, the C2 mutant, which is unable to form an intramolecular disulfide bond, has reduced binding to 14-3-3 in vitro and in vivo. These results suggest that oxidative stress may induce cell cycle arrest in part through the degradation of Cdc25C.
Savitsky
Pavel A
PA
Cardiovascular Branch, NHLBI, National Institutes of Health, Bethesda, Maryland 20892-1622, USA.
Finkel
Toren
T
D018797:Cell Cycle Proteins / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D016195:G2 Phase; D006367:HeLa Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D008938:Mitosis; D010084:Oxidation-Reduction; D010766:Phosphorylation; D011994:Recombinant Proteins / Q000378:metabolism; D020687:cdc25 Phosphatases / Q000378:metabolism*
D018797:Cell Cycle Proteins; D011994:Recombinant Proteins; D006861:Hydrogen Peroxide; C517009:CDC25C protein, human; D020687:cdc25 Phosphatases