Role of Phosphoinositide 3-Kinase Regulatory Isoforms in Development and Actin Rearrangement

Abstract

Class Ia phosphoinositide 3-kinases (PI3Ks) are heterodimers of p110 catalytic and p85 regulatory subunits that mediate a variety of cellular responses to growth and differentiation factors. Although embryonic development is not impaired in mice lacking all isoforms of the p85α gene (p85α^−/− p55α^−/− p50α^−/−) or in mice lacking the p85β gene (p85β^−/−) (D. A. Fruman, F. Mauvais-Jarvis, D. A. Pollard, C. M. Yballe, D. Brazil, R. T. Bronson, C. R. Kahn, and L. C. Cantley, Nat Genet. 26:379-382, 2000; K. Ueki, C. M. Yballe, S. M. Brachmann, D. Vicent, J. M. Watt, C. R. Kahn, and L. C. Cantley, Proc. Natl. Acad. Sci. USA 99:419-424, 2002), we show here that loss of both genes results in lethality at embryonic day 12.5 (E12.5). The phenotypes of these embryos, including subepidermal blebs flanking the neural tube at E8 and bleeding into the blebs during the turning process, are similar to defects observed in platelet-derived growth factor receptor α null (PDGFRα^−/−) mice (P. Soriano, Development 124:2691-2700, 1997), suggesting that PI3K is an essential mediator of PDGFRα signaling at this developmental stage. p85α^−/− p55α^+/+ p50α^+/+ p85β^−/− mice had similar but less severe defects, indicating that p85α and p85β have a critical and redundant function in development. Mouse embryo fibroblasts deficient in all p85α and p85β gene products (p85α^−/− p55α^−/− p50α^−/− p85β^−/−) are defective in PDGF-induced membrane ruffling. Overexpression of the Rac-specific GDP-GTP exchange factor Vav2 or reintroduction of p85α or p85β rescues the membrane ruffling defect. Surprisingly, reintroduction of p50α also restored PDGF-dependent membrane ruffling. These results indicate that class Ia PI3K is critical for PDGF-dependent actin rearrangement but that the SH3 domain and the Rho/Rac/Cdc42-interacting domain of p85, which lacks p50α, are not required for this response.

ACKNOWLEDGMENTS

S.M.B was supported by a scholarship from Boehringer Ingelheim Fonds. D.A.F was supported in part by fellowships from the Damon Runyon Cancer Research Foundation and the Leukemia and Lymphoma Society. S.M.T. was supported by the Leukemia and Lymphoma Society. This work was supported by NIH grants GM41890 and PO1-CA089021 to L.C.C, AI50831 to D.A.F, and CA75621 to S.M.T.

We thank Morris White for sharing the anti-p55γ antisera and Benjamin Neel for anti-PDGFR antibody. We also thank John Watt, Monica Kosmatka, Nicole Logsdon, and Nikki Madson for maintaining the mouse colonies.