Vesicular PtdIns(3,4,5)P3 and Rab 7 are key effectors of zygote nuclear membrane fusion

Abstract

Regulation of nuclear envelope (NE) dynamics is an important example of the universal phenomena of membrane fusion. The signalling molecules involved in nuclear membrane fusion may also be conserved in the formation of both pronuclear and zygote NEs in the fertilised egg. Here, we determine that Class I PI3-kinases are needed for in vitro nuclear envelope formation. We show that, in vivo, PtdIns(3,4,5)P3 is transiently located in vesicles around the male pronucleus at the time of nuclear envelope formation and around male and female pronuclei prior to membrane fusion. We illustrate that Class I PI3-kinase activity is also necessary for fusion of the female and male pronuclear membranes. We demonstrate, by coincidence amplified-FRET monitored by fluorescence lifetime imaging microscopy (FLIM), a protein-lipid interaction of Rab7 GTPase and PtdIns(3,4,5)P3 occurring during pronuclear membrane fusion to create the zygote nuclear envelope. We present a working model, which includes several molecular steps in the pathways controlling fusion of NE membranes.