During male pronuclei formation chromatin remodeling is uncoupled from nucleus decondensation.
Male pronucleus formation involves sperm nucleus decondensation and sperm chromatin remodeling. In sea urchins, male pronucleus decondensation was shown to be modulated by protein kinase C and a cdc2-like kinase sensitive to olomoucine in vitro assays. It was further demonstrated that olomoucine blocks SpH2B and SpH1 phosphorylation. These phosphorylations were postulated to participate in the initial steps of male chromatin remodeling during male pronucleus formation. At final steps of male chromatin remodeling, all sperm histones (SpH) disappear from male chromatin and are subsequently degraded by a cysteine protease. As a result of this remodeling, the SpH are replaced by maternal histone variants (CS). To define if sperm nucleus decondensation is coupled with sperm chromatin remodeling, we have followed the loss of SpH in zygotes treated with olomoucine. SpH degradation was followed with anti-SpH antibodies that had no cross-reactivity with CS histone variants. We found that olomoucine blocks SpH1 and SpH2B phosphorylation and inhibits male pronucleus decondensation in vivo. Interestingly, the normal schedule of SpH degradation remains unaltered in the presence of olomoucine. Taken together these results, it was concluded that male nucleus decondensation is uncoupled from the degradation of SpH associated to male chromatin remodeling. From these results, it also emerges that the phosphorylation of SpH2B and SpH1 is not required for the degradation of the SpH that is concurrent to male chromatin remodeling.
Monardes
Antonia
A
Department of Biochemistry and Molecular Biology, Universidad de Concepción, Casilla 160-C, Concepción, Chile.
Iribarren
Claudio
C
Morin
Violeta
V
Bustos
Paula
P
Puchi
Marcia
M
Imschenetzky
María
M
D000818:Animals; D002467:Cell Nucleus; D042002:Chromatin Assembly and Disassembly; D005306:Fertilization; D006657:Histones / Q000378:metabolism; D008297:Male; D010766:Phosphorylation; D012617:Sea Urchins / Q000166:cytology* / Q000196:embryology / Q000235:genetics*; D013094:Spermatozoa / Q000378:metabolism
D006657:Histones