J. Cell Biol., Volume 142, Number 4, August 24, 1998 1063-1074

Involvement of Phospholipase C1 in Mouse Egg Activation Induced by a Truncated Form of the C-kit Tyrosine Kinase Present in Spermatozoa

Claudio Sette,^* Arturo Bevilacqua, Raffaele Geremia,^* and Pellegrino Rossi^*

^* Dipartimento di Sanitá Pubblica e Biologia Cellulare, Sezione di Anatomia, Universitá di Roma Tor Vergata, Rome, Italy; and  Dipartimento di Istologia ed Embriologia Medica and Dipartimento di Psicologia, Universitá di Roma La Sapienza, Rome, Italy

Microinjection of a truncated form of the c-kit tyrosine kinase present in mouse spermatozoa (tr-kit) activates mouse eggs parthenogenetically, and tr-kit- induced egg activation is inhibited by preincubation with an inhibitor of phospholipase C (PLC) (Sette, C., A. Bevilacqua, A. Bianchini, F. Mangia, R. Geremia, and P. Rossi. 1997. Development [Camb.]. 124:2267-2274). Co-injection of glutathione-S-transferase (GST) fusion proteins containing the src-homology (SH) domains of the 1 isoform of PLC (PLC1) competitively inhibits tr-kit- induced egg activation. A GST fusion protein containing the SH3 domain of PLC1 inhibits egg activation as efficiently as the whole SH region, while a GST fusion protein containing the two SH2 domains is much less effective. A GST fusion protein containing the SH3 domain of the Grb2 adaptor protein does not inhibit tr-kit-induced egg activation, showing that the effect of the SH3 domain of PLC1 is specific. Tr-kit-induced egg activation is also suppressed by co-injection of antibodies raised against the PLC1 SH domains, but not against the PLC1 COOH-terminal region. In transfected COS cells, coexpression of PLC1 and tr-kit increases diacylglycerol and inositol phosphate production, and the phosphotyrosine content of PLC1 with respect to cells expressing PLC1 alone. These data indicate that tr-kit activates PLC1, and that the SH3 domain of PLC1 is essential for tr-kit-induced egg activation.

truncated-c-kit;  spermatozoa;  egg-activation;  phospholipase-C-1;  src-homology-domains

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