Protamine-DNA association in mammalian spermatozoa.
We have previously identified two subsets of basic nuclear proteins of mouse sperm: the protamines and a group of less basic proteins and, with the aid of a polyvalent antiserum, we have demonstrated their differential extractibility by NaCl in reducing solution (Rodman et al., J cell sci 53 (1982) 227) [9]. By affinity purification with isolated mouse sperm protamines we have obtained a protamine-specific fraction of that antiserum and a fraction that contains antibodies to the subset of less basic proteins. With those immunochemical probes we have shown the following The antigenic sites recognized by the protamine-specific antibodies are accessible, intranuclearly, only after the DNA has been removed by DNase I. The antibodies and DNA compete for binding sites on the protamines. DNA removal and consequent availability of the antigenic sites of the protamine molecules to the antibodies are possible only after displacement of the less basic proteins and chromatin decondensation have been induced. Immunoreactivity by the less basic proteins takes place without intervention of DNase. Those data indicate that the protamines are DNA-bound but that the less basic proteins are not or, alternatively, their putative DNA-binding sites do not coincide with their immuno-reactive sites. Those data also suggest that a function of the subset of less basic proteins may be to provide a shield for the protamine-DNA complex. The mouse protamine-affinity-bound antibodies are highly cross-reactive with protamines of other mammalian sperm suggesting that, despite considerable molecular diversity among mammalian protamines, the DNA-binding sites are conserved.
D000818:Animals; D001665:Binding Sites; D001666:Binding Sites, Antibody; D002843:Chromatin / Q000378:metabolism; D002846:Chromatography, Affinity; D003429:Cross Reactions; D004247:DNA / Q000378:metabolism*; D003850:Deoxyribonuclease I; D004706:Endodeoxyribonucleases; D000939:Epitopes; D006801:Humans; D008297:Male; D051379:Mice; D009687:Nuclear Proteins; D009698:Nucleoproteins / Q000378:metabolism; D011479:Protamines / Q000276:immunology / Q000378:metabolism*; D013094:Spermatozoa / Q000032:analysis*
D002843:Chromatin; D000939:Epitopes; D009687:Nuclear Proteins; D009698:Nucleoproteins; D011479:Protamines; C026588:sperm basic nuclear proteins; D004247:DNA; D004706:Endodeoxyribonucleases; D003850:Deoxyribonuclease I