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Caspase-3-mediated cleavage of ROCK I induces MLC phosphorylation and apoptotic membrane blebbing

Nature Cell Biology volume 3, pages 346–352 (2001)Cite this article

Abstract

Increased phosphorylation of myosin light chain (MLC) is necessary for the dynamic membrane blebbing that is observed at the onset of apoptosis. Here we identify ROCK I, an effector of the small GTPase Rho, as a new substrate for caspases. ROCK I is cleaved by caspase-3 at a conserved DETD1113/G sequence and its carboxy-terminal inhibitory domain is removed, resulting in deregulated and constitutive kinase activity. ROCK proteins are known to regulate MLC-phosphorylation, and apoptotic cells exhibit a gradual increase in levels of phosphorylated MLC concomitant with ROCK I cleavage. This phosphorylation, as well as membrane blebbing, is abrogated by inhibition of caspases or ROCK proteins, but both processes are independent of Rho activity. We also show that expression of active truncated ROCK I induces cell blebbing. Thus, activation of ROCK I by caspase-3 seems to be responsible for bleb formation in apoptotic cells.

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Figure 1: ROCK is cleaved by caspases during apoptosis.
Figure 2: Caspase cleavage removes a C-terminal domain from ROCK I.
Figure 3: Caspase-3 is required for cleavage of ROCK.
Figure 4: ROCK I is cleaved at Asp 1113.
Figure 5: Apoptotic cell blebbing is abrogated by inactivation of ROCK or inhibition of capases.
Figure 6: The Mr 130K kinase fragment of ROCK I binds to activated Rho but its activity is Rho-independent.
Figure 7: MLC is phosphorylated concomitantly with ROCK cleavage in a ROCK-dependent, caspase-dependent, Rho-independent manner.
Figure 8: Expression of active ROCK I induces membrane blebbing.

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Acknowledgements

We thank J. Zugaza for comments and suggestions. This work was supported by INSERM and by grants from the Association pour la Recherche sur le Cancer. M.S. is supported by a fellowship from ARERS.

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  1. INSERM U461, Faculté de Pharmacie, Châtenay-Malabry, 92296, France

    Michaël Sebbagh, Claire Renvoizé, Jocelyne Hamelin, Nicole Riché, Jacques Bertoglio & Jacqueline Bréard

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Correspondence to Jacqueline Bréard.

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Sebbagh, M., Renvoizé, C., Hamelin, J. et al. Caspase-3-mediated cleavage of ROCK I induces MLC phosphorylation and apoptotic membrane blebbing. Nat Cell Biol 3, 346–352 (2001). https://doi.org/10.1038/35070019

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