Gadd45a Expression Induces Bim Dissociation from the Cytoskeleton and Translocation to Mitochondria

Abstract

Gadd45a, a p53- and BRCA1-regulated stress protein, has been implicated in the maintenance of genomic fidelity, probably through its roles in the control of cell cycle checkpoint and apoptosis. However, the mechanism(s) by which Gadd45a is involved in the induction of apoptosis remains unclear. We show here that inducible expression of Gadd45a protein causes dissociation of Bim, a Bcl2 family member, from microtubule-associated components and translocation to mitochondria. The Bim accumulation in mitochondria enhances interaction of Bim with Bcl-2, relieves Bax from Bcl-2-bound complexes, and subsequently results in release of cytochrome c into the cytoplasm. Suppression of endogenous Bim greatly inhibits Gadd45a induction of apoptosis. Interestingly, Gadd45a interacts with elongation factor 1α (EF-1α), a microtubule-severing protein that plays an important role in maintaining cytoskeletal stability, and inhibits EF-1α-mediated microtubule bundling, indicating that the interaction of Gadd45a with EF-1α disrupts cytoskeletal stability. A mutant form of Gadd45a harboring a deletion of EF-1α-binding domain fails to inhibit microtubule stability and to induce Bim translocation to mitochondria. Furthermore, coexpression of EF-1α antagonizes Gadd45a's property of suppressing cell growth and inducing apoptosis. These findings identify a novel link that connects stress protein Gadd45a to the apoptotic machinery and address the importance of cytoskeletal stability in apoptotic response to DNA damage.

ACKNOWLEDGMENTS

This study was supported in part by the National Fundamental Research Program of China (2002CB513101), the National Natural Science Foundation of China (30225018 and 30400074), the Beijing Natural Science Foundation (7043074), and the National Institutes of Health grant R01 CA93640.

We thank Albert J. Fornace and M. Christine Hollander at NIH for providing us with Gadd45a knockout MEFs.