Assembly and characterization of nucleosomal cores on B- vs. Z-form DNA.
The ability of right- vs. left-handed alternating purine/pyrimidine copolymers to support the formation of nucleosomes has been examined by using a trout testis assembly factor. The protein, which is thermostable, has a molecular weight of 29000 and will assemble nucleosomes onto both SV40 and calf thymus DNA. This assembly factor has been used to assemble nucleosomes onto the B and Z conformations of poly[d(Gm5C)] and the B conformation of poly[d(GC)]. The isolated B-form particles, which sediment at approximately 11 S in a sucrose density gradient, contain DNA of 140-200 bases in length and the four core histones. The isolated Z-form particles, which also sediment at approximately 11 S, contain the four core histones and DNA of 170-250 bases in length. Physical analysis of the particles by absorbance and circular dichroic spectroscopy indicates that the DNA remains in the original conformation throughout the isolation procedure. Further, the particles reconstituted onto left-handed DNA compete effectively for an anti-Z DNA antibody, while the corresponding right-handed particles do not. Analytical sedimentation velocity determinations indicate that the B-form poly[d(Gm5C)] and poly[d(GC)] particles sediment at 11.2 and 11.1 S, respectively. In contrast, the poly[d(Gm5C)] Z-form particles have an S20,w of 10.6 S. The differences in the sedimentation velocity and the density of the cores, and in the lengths of DNA associated with the particles, suggest that the conformation of the DNA affects the manner in which it associates with the histone octamer.
Miller
F D
FD
Dixon
G H
GH
Rattner
J B
JB
van de Sande
J H
JH
D000818:Animals; D004247:DNA / Q000302:isolation & purification / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism; D006657:Histones / Q000378:metabolism*; D008297:Male; D008854:Microscopy, Electron; D009690:Nucleic Acid Conformation; D009707:Nucleosomes / Q000378:metabolism / Q000648:ultrastructure*; D011089:Polydeoxyribonucleotides; D013539:Simian virus 40; D013056:Spectrophotometry, Ultraviolet; D013735:Testicular Hormones / Q000378:metabolism; D013737:Testis / Q000378:metabolism / Q000648:ultrastructure*; D014337:Trout
D004279:DNA, Viral; D006657:Histones; D009707:Nucleosomes; D011089:Polydeoxyribonucleotides; D013735:Testicular Hormones; C019530:poly(dC-dG); D004247:DNA