Role of individual histone tyrosines in the formation of the nucleosome complex.
We have determined the accessibility of histone tyrosine residues to react with p-nitrobenzenesulfonyl fluoride (NBSF) in intact nuclei, salt-dissociated nucleosomes, isolated histone complexes, and individual core histones. Of the 15 core histone tyrosine residues, 13 are inaccessible in native nucleosomes; only Tyr121 near the C-terminus of H2B is fully accessible, and Tyr54 of H3 is partially accessible under near-physiological conditions. When H1 and the basic N-terminal tails of the core histones are dissociated from the DNA by treating nuclei with 0.4 and 0.8 M NaCl, the two tyrosines which are adjacent to the basic regions of H2B and H3 become accessible as well. This indicates that these tyrosine residues may be involved in histone-DNA interactions, either directly or indirectly. When the H2A-H2B dimers are dissociated from the chromatin by raising the NaCl concentration to 1.2 M, three to four tyrosines located in the structured regions of H2B and H4 are exposed, suggesting that these tyrosine residues may be located at the dimer-tetramer interface. Dissociating all the histones from the DNA at an even higher ionic strength as a mixture of dimers, tetramers, and octamers does not change the pattern of Tyr exposure, but reduces the reactivity of the tyrosines at the dimer-tetramer interface as would be expected from the reassociation of H2A-H2B dimers and H3-H4 tetramers.(ABSTRACT TRUNCATED AT 250 WORDS)
Zweidler
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Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111.
D000595:Amino Acid Sequence; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006657:Histones / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D007202:Indicators and Reagents; D007700:Kinetics; D046911:Macromolecular Substances; D008958:Models, Molecular; D009578:Nitrobenzenes / Q000494:pharmacology; D009707:Nucleosomes / Q000378:metabolism*; D009994:Osmolar Concentration; D010449:Peptide Mapping; D011487:Protein Conformation; D051381:Rats; D013950:Thymus Gland / Q000378:metabolism; D014443:Tyrosine
D006657:Histones; D007202:Indicators and Reagents; D046911:Macromolecular Substances; D009578:Nitrobenzenes; D009707:Nucleosomes; C034442:4-nitrobenzenesulfonyl fluoride; D014443:Tyrosine