Myristoylated alanine-rich C kinase substrate, but not Ca2+/calmodulin-dependent protein kinase II, is the mediator in cortical granules exocytosis.
Sperm-egg fusion induces cortical granules exocytosis (CGE), a process that ensures the block to polyspermy. CGE can be induced independently by either a rise in intracellular calcium concentration or protein kinase C (PKC) activation. We have previously shown that myristoylated alanine-rich C kinase substrate (MARCKS) cross-links filamentous actin (F-actin) and regulates its reorganization. This activity is reduced either by PKC-induced MARCKS phosphorylation (PKC pathway) or by its direct binding to calmodulin (CaM; CaM pathway), both inducing MARCKS translocation, F-actin reorganization, and CGE. Currently, we examine the involvement of Ca(2)(+)/CaM-dependent protein kinase II (CaMKII) and MARCKS in promoting CGE and show that PKC pathway can compensate for lack of Ca(2)(+)/CaM pathway. Microinjecting eggs with either overexpressed protein or complementary RNA of constitutively active alphaCaMKII triggered resumption of second meiotic division, but induced CGE of an insignificant magnitude compared with CGE induced by wt alphaCaMKII. Microinjecting eggs with mutant-unphosphorylatable MARCKS reduced the intensity of 12-O-tetradecanoylphorbol 13-acetate or ionomycin-induced CGE by 50%, indicating that phosphorylation of MARCKS by novel and/or conventional PKCs (n/cPKCs) is a pivotal event associated with CGE. Moreover, we were able to demonstrate cPKCs involvement in ionomycin-induced MARCKS translocation and CGE. These results led us to propose that MARCKS, rather than CaMKII, as a key mediator of CGE.
Tsaadon
Lina
L
Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel-Aviv University, Ramat Aviv, Tel-Aviv 69978, Israel.
Kaplan-Kraicer
Ruth
R
Shalgi
Ruth
R
D000818:Animals; D001483:Base Sequence; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D002970:Cleavage Stage, Ovum / Q000502:physiology*; D005089:Exocytosis; D005260:Female; D005818:Genetic Engineering; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000502:physiology*; D015759:Ionomycin / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology*; D008845:Microinjections; D008969:Molecular Sequence Data; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; D010766:Phosphorylation; D012333:RNA, Messenger / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D014178:Translocation, Genetic / Q000187:drug effects
D047908:Intracellular Signaling Peptides and Proteins; D007476:Ionophores; C000619712:Marcks protein, rat; D008565:Membrane Proteins; D012333:RNA, Messenger; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; D015759:Ionomycin; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2