Protein kinase C-mediated interphase lamin B phosphorylation and solubilization.
Disassembly of the sperm nuclear envelope at fertilization is one of the earliest events in the development of the male pronucleus. We report that nuclear lamina disassembly in interphase sea urchin egg cytosol is a result of lamin B phosphorylation mediated by protein kinase C (PKC). Lamin B of permeabilized sea urchin sperm nuclei incubated in fertilized egg G1 phase cytosolic extract is phosphorylated within 1 min of incubation and solubilized prior to sperm chromatin decondensation. Phosphorylation is Ca2+-dependent. It is reversibly inhibited by the PKC-specific inhibitor chelerythrine, a PKC pseudosubstrate inhibitor peptide, and a PKC substrate peptide, but not by inhibitors of PKA, p34(cdc2) or calmodulin kinase II. Phosphorylation is inhibited by immunodepletion of cytosolic PKC and restored by addition of purified rat brain PKC. Sperm lamin B is a substrate for rat brain PKC in vitro, resulting in lamin B solubilization. Two-dimensional phosphopeptide maps of lamin B phosphorylated by the cytosolic kinase and by purified rat PKC are virtually identical. These data suggest that PKC is the major kinase required for interphase disassembly of the sperm lamina.
Collas
P
P
Department of Biochemistry, Norwegian College of Veterinary Medicine, 0033 Oslo, Norway. philippe.collas@veths.no
Thompson
L
L
Fields
A P
AP
Poccia
D L
DL
Courvalin
J C
JC
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002451:Cell Compartmentation; D002467:Cell Nucleus / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005306:Fertilization; D007399:Interphase; D034921:Lamin Type B; D034882:Lamins; D008297:Male; D009687:Nuclear Proteins / Q000378:metabolism*; D010063:Ovum / Q000378:metabolism; D010449:Peptide Mapping; D010748:Phosphopeptides / Q000737:chemistry; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D051381:Rats; D012617:Sea Urchins; D013094:Spermatozoa / Q000378:metabolism
D004791:Enzyme Inhibitors; D034921:Lamin Type B; D034882:Lamins; D009687:Nuclear Proteins; D010748:Phosphopeptides; D011493:Protein Kinase C; D002118:Calcium