Identification of a starfish egg PLC-gamma that regulates Ca2+ release at fertilization.

TitleIdentification of a starfish egg PLC-gamma that regulates Ca2+ release at fertilization.
Publication TypeJournal Article
Year of Publication2004
AuthorsRunft LL, Carroll DJ, Gillett J, Giusti AF, O'Neill FJ, Foltz KR
JournalDev Biol
Volume269
Issue1
Pagination220-36
Date Published2004 May 1
ISSN0012-1606
KeywordsAmino Acid Sequence, Animals, Calcium, Fertilization, Molecular Sequence Data, Ovum, Phospholipase C gamma, src Homology Domains, Starfish, Type C Phospholipases
Abstract

At fertilization, eggs undergo a cytoplasmic free Ca2+ rise, which is necessary for stimulating embryogenesis. In starfish eggs, studies using inhibitors designed against vertebrate proteins have shown that this Ca2+ rise requires an egg Src family kinase (SFK) that directly or indirectly activates phospholipase C-gamma (PLC-gamma) to produce IP3, which triggers Ca2+ release from the egg's endoplasmic reticulum (ER) [reviewed in Semin. Cell Dev. Biol. 12 (2001) 45]. To examine in more detail the endogenous factors in starfish eggs that are required for Ca2+ release at fertilization, an oocyte cDNA encoding PLC-gamma was isolated from the starfish Asterina miniata. This cDNA, designated AmPLC-gamma, encodes a protein with 49% identity to mammalian PLC-gamma1. A 58-kDa Src family kinase interacted with recombinant AmPLC-gamma Src homology 2 (SH2) domains in a specific, fertilization-responsive manner. Immunoprecipitations of sea urchin egg PLC-gamma using an affinity-purified antibody directed against AmPLC-gamma revealed fertilization-dependent phosphorylation of PLC-gamma. Injecting starfish eggs with the tandem SH2 domains of AmPLC-gamma (which inhibits PLC-gamma activation) specifically inhibited Ca2+ release at fertilization. These results indicate that an endogenous starfish egg PLC-gamma interacts with an egg SFK and mediates Ca2+ release at fertilization via a PLC-gamma SH2 domain-mediated mechanism.

DOI10.1016/j.ydbio.2004.01.031
Alternate JournalDev. Biol.
PubMed ID15081369