Binding of benzo[a]pyrene to histones and altered affinity of modified histone 1 for deoxyribonucleic acid.
The covalent binding of benzo[a]pyrene (B[a]P) to acid extractable chromosomal proteins and the subsequent effect on histone 1-DNA interaction have been characterized in a model system by utilizing calf thymus nuclei as targets and rat liver microsomes as an exogenous source of enzymes for the metabolic activation of B[a]P. A two-step ion-exchange chromatography and desalting procedure was employed for removing noncovalently bound B[a]P and other contaminants. Fluorography of acetic acid-urea and Triton-acetic acid-urea-polyacrylamide gels indicated that H1 and H3 were the only principal histone targets in [3H]B[a]P-modified calf thymus nuclei. The validity of this assignment was confirmed by comparison of the chromatographic distributions of [3H]B[a]P cpm among peptides derived from the HClO4- soluble (H1) and HClO4-insoluble (core histones) protein fractions to the distributions obtained for authentic individual histone fractions. Comparison of amino acid compositions in individual peptide fractions which bound [3H]B[a]P differentially yielded some insight into the probable target amino acid residues for B[a]P binding. On the basis of electrophoresis in polyacrylamide gels, it appeared as if B[a]P had bound to multiple subfractions of H1 and H3. The equivalent distribution of covalently attached [3H]B[a]P among the major peptides of H1 and H3 modified either in intact nuclei or while free in solution implied that the relative accessibility of major portions of the H1 and H3 molecules for covalent B[a]P binding is not affected by interactions with DNA or other chromosomal proteins. Covalent attachment of [3H]B[a]P to purified H1 reduced the affinity of this histone for DNA-cellulose.
D000818:Animals; D001564:Benzo(a)pyrene; D001580:Benzopyrenes / Q000378:metabolism*; D001665:Binding Sites; D001711:Biotransformation; D002417:Cattle; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D006657:Histones / Q000378:metabolism*; D066298:In Vitro Techniques; D008862:Microsomes, Liver / Q000378:metabolism; D051381:Rats; D013950:Thymus Gland / Q000378:metabolism
D001580:Benzopyrenes; D006657:Histones; D001564:Benzo(a)pyrene; D004247:DNA