Spy1 expression prevents normal cellular responses to DNA damage: inhibition of apoptosis and checkpoint activation.
Spy1 is the originally identified member of the Speedy/Ringo family of vertebrate cell cycle regulators, which can control cell proliferation and survival through the atypical activation of cyclin-dependent kinases. Here we report a role for Spy1 in apoptosis and checkpoint activation in response to UV irradiation. Using an inducible system allowing for regulated expression of Spy1, we show that Spy1 expression prevents activation of caspase-3 and suppresses apoptosis in response to UV irradiation. Spy1 expression also allows for UV irradiation-resistant DNA synthesis and permits cells to progress into mitosis, as demonstrated by phosphorylation on histone H3, indicating that Spy1 expression can inhibit the S-phase/replication and G2/M checkpoints. We demonstrate that Spy1 expression inhibits phosphorylation of Chk1, RPA, and histone H2A.X, which may directly contribute to the decrease in apoptosis and checkpoint bypass. Furthermore, mutation of the conserved Speedy/Ringo box, known to mediate interaction with CDK2, abrogates the ability of Spy1 to inhibit apoptosis and the phosphorylation of Chk1 and RPA. The data presented indicate that Spy1 expression allows cells to evade checkpoints and apoptosis and suggest that Spy1 regulation of CDK2 is important for the response to DNA damage.
Gastwirt
Randy F
RF
Biomedical Sciences Program, Department of Chemistry and Biochemistry, and Moores University of California San Diego Cancer Center, University of California San Diego, La Jolla, California 92093-0367, USA.
Slavin
Daniela A
DA
McAndrew
Christopher W
CW
Donoghue
Daniel J
DJ
D017209:Apoptosis / Q000502:physiology*; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D051357:Cyclin-Dependent Kinase 2 / Q000378:metabolism; D004249:DNA Damage / Q000502:physiology*; D004789:Enzyme Activation; D005786:Gene Expression Regulation; D006801:Humans; D009154:Mutation; D010011:Osteocytes / Q000166:cytology; D015398:Signal Transduction
D018797:Cell Cycle Proteins; C486530:SPDYA protein, human; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2; D053148:Caspase 3