Proteolytic cleavage of ras GTPase-activating protein during apoptosis.
p120-ras GTPase-activating protein (rasGAP) associates with Ras and negatively regulates Ras signaling by stimulating the intrinsic rate of Ras GTPase activity. rasGAP also associates with other cellular signaling proteins which suggest that rasGAP may play a role in coordinating other signal transduction pathways. Disruption of rasGAP in vivo results in extensive apoptosis. Fas-mediated apoptosis results in the activation of caspases that cleave cellular substrates which are important for maintaining cytoplasmic and nuclear integrity. We show here that rasGAP is proteolytically cleaved by caspases early in Fas-induced apoptosis of Jurkat cells. rasGAP was also cleaved by DNA-damaging chemotherapeutic agents and TNF-related apoptosis inducing ligand (TRAIL), also known as Apo2L. Based on the size of the products generated by cleavage of deletion mutants of rasGAP we predict that cleavage of rasGAP occurs in the hydrophobic region and between the SH2(2) and ras-p21 interacting domain which would leave an intact ras-p21 interacting domain. Interestingly, cleavage of rasGAP in vitro enhanced rasGAP hydrolysis activity. Our results demonstrate that diverse apoptotic stimuli cause caspase-mediated cleavage of rasGAP early in apoptosis.
Wen
L P
LP
Department of Pulmonary and Critical Care Medicine, Stanford University, Stanford, California 94305-5236, USA.
Madani
K
K
Martin
G A
GA
Rosen
G D
GD
D017209:Apoptosis; D001665:Binding Sites; D053148:Caspase 3; D053179:Caspase 7; D020169:Caspases / Q000378:metabolism*; D020690:GTPase-Activating Proteins; D006801:Humans; D006868:Hydrolysis; D019169:Jurkat Cells; D010449:Peptide Mapping; D011506:Proteins / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D019014:fas Receptor / Q000378:metabolism*; D020703:ras GTPase-Activating Proteins
D020690:GTPase-Activating Proteins; D011506:Proteins; D019014:fas Receptor; D020703:ras GTPase-Activating Proteins; C505407:CASP3 protein, human; C505520:CASP7 protein, human; D053148:Caspase 3; D053179:Caspase 7; D020169:Caspases