Mouse cPLA2gamma, a novel oocyte and early embryo-abundant phospholipase A2 gamma-like protein, is targeted to the nuclear envelope during germinal vesicle breakdown.
This report documents the characterization of a novel mouse oocyte protein which was originally identified by microsequence analysis of a 67.8 kDa protein spot (pI 5.7) on a Coomassie-stained two-dimensional (2D) gel of murine egg proteins. Tandem mass spectroscopic analysis of the peptides obtained from the cored protein yielded sequences that appeared to match only ovary, egg, and preimplantation embryo cDNAs. We then cloned the novel gene by RACE-PCR, and analysis of the deduced cDNA sequence found that this maternal product was approximately 56% identical to human cytosolic phospholipase A2gamma (cPLA2gamma). Based on this sequence homology, we named the molecule mouse cytosolic phospholipase A2gamma (cPLA2gamma). As with human cPLA2gamma, mouse cPLA2gamma contains a lipase consensus sequence and lacks the calcium binding domain that is found in other PLA2 proteins. However, mouse cPLA2gamma is different from human cPLA2gamma in that mouse cPLA2gamma expression is restricted to the ovary and that the protein does not contain the myristoylation and prenylation lipid-anchoring motifs that are present in human cPLA2gamma. Within oocytes, mouse cPLA2gamma localizes mainly to the oocyte cortex and to the nucleoplasm. Interestingly, during germinal vesicle breakdown, mouse cPLA2gamma aggregates dynamically relocate from the oocyte cortex to the nuclear envelope, suggesting a possible role for this putative egg-restricted phospholipase A2gamma in membrane remodeling. Furthermore, mouse cPLA2gamma protein continues to be expressed in the embryo until the 4-8-cell stage of development, suggesting that mouse cPLA2gamma may function as a previously uncharacterized maternal effect gene.
Vitale
Alejandra
A
Weill Medical College of Cornell University, New York, NY 10021, USA.
Perlin
Julie
J
Leonelli
Lauriebeth
L
Herr
John
J
Wright
Paul
P
Digilio
Laura
L
Coonrod
Scott
S
D020816:Amino Acid Motifs / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002970:Cleavage Stage, Ovum / Q000378:metabolism*; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D054513:Group IV Phospholipases A2; D007150:Immunohistochemistry; D013058:Mass Spectrometry; D051379:Mice; D008969:Molecular Sequence Data; D009685:Nuclear Envelope / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism*; D010053:Ovary / Q000033:anatomy & histology / Q000378:metabolism; D010741:Phospholipases A / Q000235:genetics* / Q000378:metabolism*; D054467:Phospholipases A2; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017385:Sequence Homology
D017931:DNA Primers; D018076:DNA, Complementary; D010741:Phospholipases A; D054513:Group IV Phospholipases A2; D054467:Phospholipases A2