Phosphorylation protects sperm-specific histones H1 and H2B from proteolysis after fertilization.
At intermediate stages of male pronucleus formation, sperm-derived chromatin is composed of hybrid nucleoprotein particles formed by sperm H1 (SpH1), dimers of sperm H2A-H2B (SpH2A-SpH2B), and a subset of maternal cleavage stage (CS) histone variants. At this stage in vivo, the CS histone variants are poly(ADP-ribosylated), while SpH2B and SpH1 are phosphorylated. We have postulated previously that the final steps of sperm chromatin remodeling involve a cysteine-protease (SpH-protease) that degrades sperm histones in a specific manner, leaving the maternal CS histone variants unaffected. More recently we have reported that the protection of CS histones from degradation is determined by the poly(ADP-ribose) moiety of these proteins. Because of the selectivity displayed by the SpH-protease, the coexistence of a subset of SpH together with CS histone variants at intermediate stages of male pronucleus remodeling remains intriguing. Consequently, we have investigated the phosphorylation state of SpH1 and SpH2B in relation to the possible protection of these proteins from proteolytic degradation. Histones H1 and H2B were purified from sperm, phosphorylated in vitro using the recombinant alpha-subunit of casein kinase 2, and then used as substrates in the standard assay of the SpH-protease. The phosphorylated forms of SpH1 and SpH2B were found to remain unaltered, while the nonphosphorylated forms were degraded. On the basis of this result, we postulate a novel role for the phosphorylation of SpH1 and SpH2B that occurs in vivo after fertilization, namely to protect these histones against degradation at intermediate stages of male chromatin remodeling.
Morin
V
V
Department of Molecular Biology, Universidad de Concepci¿on, Casilla 160-C, Concepci¿on, Chile.
Acuña
P
P
Díaz
F
F
Inostroza
D
D
Martinez
J
J
Montecino
M
M
Puchi
M
M
Imschenetzky
M
M
D000595:Amino Acid Sequence; D000818:Animals; D002843:Chromatin / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism; D005260:Female; D005306:Fertilization / Q000502:physiology*; D006657:Histones / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D066298:In Vitro Techniques; D008297:Male; D010766:Phosphorylation; D012617:Sea Urchins; D013094:Spermatozoa / Q000378:metabolism*
D002843:Chromatin; D006657:Histones; D010450:Endopeptidases