Factors affecting nucleosome disassembly by protamines in vitro. Histone hyperacetylation and chromatin structure, time dependence, and the size of the sperm nuclear proteins.
Histone displaced in vitro from nuclei by protamine competition display a higher degree of hyperacetylation than the residual histones. In addition, hyperacetylated core particle pools are disassembled in vitro with a higher efficiency than control or nonacetylated core particles and when analyzed by electron microscopy display an elongated shape (length/width ratio = 1.52 +/- 0.19) instead of the round compact shape of control nucleosomes (length/width ratio = 1.06 +/- 0.06). In the absence of histone hyperacetylation, the fish protamines, salmine and iridine (32-33 residues), are relatively inefficient in disassembling nucleosomal core particles in vitro as compared to the large (65-70 residues), tyrosine-containing protamines from rooster (galline), squid, and cuttlefish which disassemble nucleosomes in a range of protamine concentrations close to physiological. The fact that an artificially cross-linked salmine dimer acquires the ability of the large protamines from rooster, squid, and cuttlefish to disassemble core particles in vitro and also binds more tightly to the DNA, suggests that the size of the sperm nuclear protamines is a critical factor in this process. Even when the core histones of spermatid chromatin are hyperacetylated in the trout testis, the replacement process by iridine or salmine is slow and time-dependent in vitro. However, since spermiogenesis in trout occurs over several weeks, the slow in vitro nucleosome disassembly process by salmine is sufficient to allow complete displacement, thus supporting the hypothesis that a protamine-mediated displacement of the histones from DNA in vivo may take place in the salmonid fishes by a mechanism similar to that in the rooster, squid, and cuttlefish.
Oliva
R
R
Department of Medical Biochemistry, Faculty of Medicine, University of Calgary, Alberta, Canada.
Bazett-Jones
D
D
Mezquita
C
C
Dixon
G H
GH
D000107:Acetylation; D000818:Animals; D002645:Chickens; D002843:Chromatin / Q000648:ultrastructure*; D005399:Fishes; D006367:HeLa Cells / Q000032:analysis; D006657:Histones / Q000378:metabolism*; D008297:Male; D008854:Microscopy, Electron; D009698:Nucleoproteins / Q000378:metabolism; D011479:Protamines / Q000378:metabolism*; D012473:Salmine / Q000378:metabolism*; D013091:Spermatogenesis; D013094:Spermatozoa / Q000032:analysis*
D002843:Chromatin; D006657:Histones; D009698:Nucleoproteins; D011479:Protamines; C001472:iridine; D012473:Salmine