The reconstitution of a hybrid histone octamer containing avian 110Cys-des-thio-histone H3 and sea-urchin 73Cys-histone H4.
A hybrid histone octamer was reconstituted from erythrocyte H2A and H2B, avian [110 Cys-des-thio]histone H3 and the sea-urchin sperm [73Cys]H4 variant. [110Cys-Des-thio]histone H3 was prepared by reaction of natural H3 with Raney nickel. The ability of the hybrid octamer to crystallize to the same form as the natural octamer demonstrated that the chemical modification of cysteine to alanine in H3 and the mutation from threonine to cysteine in sperm H4 do not alter histone-histone interactions in the octamer. Since the sulfhydryl groups of both H4 molecules are fully accessible to 5,5'-dithiobis(2-nitrobenzoate) these residues provide suitable sites for the introduction of a single cysteine-specific label per H4 molecule in the octamer.
Greyling
H J
HJ
Department of Biochemistry, University of Cape Town, Rondebosch, South Africa.
Sewell
B T
BT
Von Holt
C
C
D000596:Amino Acids / Q000302:isolation & purification; D000818:Animals; D002645:Chickens; D004912:Erythrocytes / Q000378:metabolism; D006657:Histones / Q000302:isolation & purification*; D006824:Hybridization, Genetic; D008297:Male; D012617:Sea Urchins; D013094:Spermatozoa / Q000378:metabolism
D000596:Amino Acids; D006657:Histones