Yeast histone H3 and H4 amino termini are important for nucleosome assembly in vivo and in vitro: redundant and position-independent functions in assembly but not in gene regulation.
The hydrophilic amino-terminal sequences of histones H3 and H4 extend from the highly structured nucleosome core. Here we examine the importance of the amino termini and their position in the nucleosome with regard to both nucleosome assembly and gene regulation. Despite previous conclusions based on nonphysiological nucleosome reconstitution experiments, we find that the histone amino termini are important for nucleosome assembly in vivo and in vitro. Deletion of both tails, a lethal event, alters micrococcal nuclease-generated nucleosomal ladders, plasmid superhelicity in whole cells, and nucleosome assembly in cell extracts. The H3 and H4 amino-terminal tails have redundant functions in this regard because the presence of either tail allows assembly and cellular viability. Moreover, the tails need not be attached to their native carboxy-terminal core. Their exchange re-establishes both cellular viability and nucleosome assembly. In contrast, the regulation of GAL1 and the silent mating loci by the H3 and H4 tails is highly disrupted by exchange of the histone amino termini.
Ling
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X
Department of Biological Chemistry, University of California at Los Angeles (UCLA) School of Medicine, 90095, USA.
Harkness
T A
TA
Schultz
M C
MC
Fisher-Adams
G
G
Grunstein
M
M
D001483:Base Sequence; D002843:Chromatin / Q000378:metabolism; D017931:DNA Primers; D004278:DNA, Superhelical / Q000737:chemistry; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D016195:G2 Phase; D005690:Galactose / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D006657:Histones / Q000737:chemistry* / Q000502:physiology*; D008836:Micrococcal Nuclease / Q000378:metabolism; D008969:Molecular Sequence Data; D009707:Nucleosomes / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D014158:Transcription, Genetic
D002843:Chromatin; D017931:DNA Primers; D004278:DNA, Superhelical; D005656:Fungal Proteins; D006657:Histones; D009707:Nucleosomes; D011994:Recombinant Proteins; D008836:Micrococcal Nuclease; D005690:Galactose