Two kinase activities are sufficient for sea urchin sperm chromatin decondensation in vitro.
Decondensation of compact and inactive sperm chromatin by egg cytoplasm at fertilization is necessary to convert the male germ cell chromatin to an active somatic form. We studied decondensation of sea urchin sperm nuclei in a cell-free extract of sea urchin eggs to define conditions promoting decondensation. We find that egg cytosol specifically phosphorylates two sperm-specific (Sp) histones in vitro in the same regions as in vivo. This activity is blocked by olomoucine, an inhibitor of cdc2-like kinases, but not by chelerythrine, an inhibitor of protein kinase C (PKC). PKC phosphorylates and solubilizes the sperm nuclear lamina, one requirement for decondensation. Olomoucine, which does not inhibit lamina removal, blocks sperm nuclear decondensation in the same concentration range over which it is effective in blocking Sp histone phosphorylation. In a system free of other soluble proteins, neither PKC nor cdc2 alone elicit sperm chromatin decondensation, but the two act synergistically to decondense sperm nuclei. We conclude that two kinases activities are sufficient for sea urchin male pronuclear decondensation in vitro, a lamin kinase (PKC) and a cdc2-like Sp histone kinase.
Department of Biology, Amherst College, Amherst, Massachusetts 01002, USA.
S
S
Stephens
B
B
Beyer
U
U
Balthazar-Stablein
R
R
Duncan
M
M
Kostacos
M
M
Lukoma
G R
GR
Green
D
D
Poccia
D000470:Alkaloids; D000818:Animals; D053119:Benzophenanthridines; D016203:CDC2 Protein Kinase / Q000737:chemistry / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism*; D004795:Enzyme Stability; D006657:Histones / Q000378:metabolism; D007701:Kinetin; D008297:Male; D010063:Ovum / Q000378:metabolism; D010617:Phenanthridines / Q000378:metabolism; D010766:Phosphorylation; D011493:Protein Kinase C / Q000737:chemistry / Q000378:metabolism*; D011687:Purines / Q000378:metabolism; D012617:Sea Urchins / Q000201:enzymology* / Q000235:genetics; D013094:Spermatozoa / Q000166:cytology / Q000201:enzymology*
D000470:Alkaloids; D053119:Benzophenanthridines; D002843:Chromatin; D006657:Histones; D010617:Phenanthridines; D011687:Purines; C090046:olomoucine; C016299:chelerythrine; D011493:Protein Kinase C; D016203:CDC2 Protein Kinase; D007701:Kinetin