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F-actin-binding domain of c-Abl regulates localized phosphorylation of C3G: role of C3G in c-Abl-mediated cell death

Oncogene volume 29, pages 4528–4542 (2010)Cite this article

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Abstract

The c-Abl tyrosine kinase maintains cellular homeostasis through its ability to regulate apoptosis and actin dynamics. In vivo, c-Abl activity is stringently regulated and mechanisms involved are not fully understood. Here, we identified the Rap1 guanine nucleotide exchange factor, C3G (RapGEF1), as a substrate and an effector of c-Abl-mediated functions. Ectopic expression of c-Abl in mammalian cell lines, known to induce apoptosis, resulted in phosphorylation of endogenous C3G on Y504 coincident with cell detachment and chromatin condensation. Phosphorylation of C3G coincided with restricted c-Abl activation in regions rich in actin, and was dependent on cellular F-actin dynamics. Unlike C3G or c-Abl, p-C3G was resistant to detergent extraction, suggesting its enhanced affinity for the cytoskeleton. Localized C3G phosphorylation and coincidence with cells undergoing cell death was dependent on F-actin-binding domain (FABD) of c-Abl. Activation of endogenous c-Abl by oxidative stress was associated with phosphorylation of cellular C3G on Y504. Inhibition of C3G expression and function using RNAi or dominant-negative approaches inhibited c-Abl-mediated cell death. These findings identify C3G as a novel target of c-Abl and also show that FABD of c-Abl is essential for regulation of its restricted activation to induce apoptosis.

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Acknowledgements

We thank Drs PJS Stork (Oregon Health and Science University), Richard Van Etten (Tufts University), Patrick Casey (Duke University), S Tanaka (Hokkaido University School of Medicine), Bruce Mayer (University of Connecticut Health Center) for gifts of various constructs. We thank Dr Ghanshyam Swarup for support extended and for critically reading the paper. Assistance received from Ms Nandini Rangaraj during use of confocal microscopes is gratefully acknowledged. AM was supported by a fellowship from the Council for Scientific and Industrial Research. This work was supported by the Department of Science and Technology, Government of India through a grant awarded to VR.

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  1. Centre for Cellular and Molecular Biology, Council of Scientific and Industrial Research (CSIR), Uppal Road, Hyderabad, India

    A Mitra & V Radha

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Correspondence to V Radha.

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Mitra, A., Radha, V. F-actin-binding domain of c-Abl regulates localized phosphorylation of C3G: role of C3G in c-Abl-mediated cell death. Oncogene 29, 4528–4542 (2010). https://doi.org/10.1038/onc.2010.113

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