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Originally published In Press as doi:10.1074/jbc.M001091200 on March 22, 2000

J. Biol. Chem., Vol. 275, Issue 22, 16788-16794, June 2, 2000
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Evidence That Fertilization Activates Starfish Eggs by Sequential Activation of a Src-like Kinase and Phospholipase Cgamma *

Andrew F. GiustiDagger §, Wenqing Xu, Beth HinkleDagger §, Mark TerasakiDagger §, and Laurinda A. JaffeDagger §

From the Dagger  Marine Biological Laboratory, Woods Hole, Massachusetts 02543, the § Department of Physiology, University of Connecticut Health Center, Farmington, Connecticut 06032, and the  Department of Biological Structure, Biomolecular Structure Center, University of Washington, Seattle, Washington 98195

Recent evidence has indicated a requirement for a Src family kinase in initiating Ca2+ release at fertilization in starfish eggs (Giusti, A. F., Carroll, D. J., Abassi, Y. A., Terasaki, M., Foltz, K. R., and Jaffe, L. A. (1999) J. Biol. Chem. 274, 29318-29322). We now show that injection of Src protein into starfish eggs initiates Ca2+ release and DNA synthesis, as occur at fertilization. These responses depend on the phosphorylation state of the Src protein; only the kinase active form is effective. Like Ca2+ release at fertilization, the Ca2+ release in response to Src protein injection is inhibited by prior injection of the SH2 domains of phospholipase Cgamma . These findings support the conclusion that in starfish, sperm-egg interaction causes egg activation by sequential activation of a Src-like kinase and phospholipase Cgamma . Injection of the SH2 domain of Src, which inhibits Ca2+ release at fertilization, does not inhibit Ca2+ release caused by Src protein injection. This indicates that the requirement for a Src SH2 domain interaction is upstream of Src activation in the pathway leading to Ca2+ release at fertilization.

* This work was supported by grants from the National Institutes of Health (to A. G. and L. A. J.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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