Caspase-3 and caspase-7 but not caspase-6 cleave Gas2 in vitro: implications for microfilament reorganization during apoptosis.
Apoptosis is characterized by proteolysis of specific cellular proteins by a family of cystein proteases known as caspases. Gas2, a component of the microfilament system, is cleaved during apoptosis and the cleaved form specifically regulates microfilaments and cell shape changes. We now demonstrate that Gas2 is a substrate of caspase-3 but not of caspase-6. Proteolytic processing both in vitro and in vivo is dependent on aspartic residue 279. Gas2 cleavage was only partially impaired in apoptotic MCF-7 cells which lack caspase-3, thus indicating that different caspases can process Gas2 in vivo. In vitro Gas2 was processed, albeit with low affinity, by caspase-7 thus suggesting that this caspase could be responsible for the incomplete Gas2 processing observed in UV treated MCF-7 cells. In vivo proteolysis of Gas2 was detected at an early stage of the apoptotic process when the cells are still adherent on the substrate and it was coupled to the specific rearrangement of the microfilament characterizing cell death. Finally we also demonstrated that Gas2 in vitro binds to F-actin, but this interaction was unaffected by the caspase-3 dependent proteolytic processing.
Sgorbissa
A
A
Dipartimento di Scienze e Tecnologie Biomediche, Sezione di Biologia, Universita' di Udine, p.le Kolbe 4, Italy.
Benetti
R
R
Marzinotto
S
S
Schneider
C
C
Brancolini
C
C
D016475:3T3 Cells; D008841:Actin Cytoskeleton / Q000502:physiology* / Q000648:ultrastructure; D000199:Actins / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001943:Breast Neoplasms; D019556:COS Cells; D053148:Caspase 3; D053178:Caspase 6; D053179:Caspase 7; D020169:Caspases / Q000378:metabolism*; D005260:Female; D006801:Humans; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism*; D008954:Models, Biological; D011994:Recombinant Proteins / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000199:Actins; C490936:GAS2 protein, human; C099165:Gas2 protein, mouse; D008840:Microfilament Proteins; D011994:Recombinant Proteins; C505407:CASP3 protein, human; C505514:CASP6 protein, human; C505520:CASP7 protein, human; C505409:Casp3 protein, mouse; C505516:Casp6 protein, mouse; C505524:Casp7 protein, mouse; D053148:Caspase 3; D053178:Caspase 6; D053179:Caspase 7; D020169:Caspases