title: Bipartite structure of the proximal promoter of a human H4 histone gene.
abstract: The proximal promoter of the human H4 histone gene FO108 contains two regions of in vivo protein-DNA interaction, Sites I and II. Electrophoretic mobility shift assays using a radiolabeled DNA probe revealed that several proteins present in HeLa cell nuclear extracts bound specifically to Site I (nt-125 to nt-86). The most prominent complex, designated HiNF-C, and a complex of greater mobility, HiNF-C', were specifically compatable by an Sp1 consensus oligonucleotide. Fractionation of HiNF-C using wheat germ agglutinin affinity chromatography suggested that, like Sp1, HiNF-C contains N-acetylglucosamine moieties. Two minor complexes of even greater mobility, designated HiNF-E and F, were compatable by ATF consensus oligonucleotides. A DNA probe carrying a site-specific mutation in the distal portion of Site I failed to bind HiNF-E, indicating that this protein associated specifically to this region. UV cross-linking analysis showed that several proteins of different molecular weights interact specifically with Site I. These data indicate that Site I possesses a bipartite structure and that multiple proteins present in HeLa cell nuclear extracts interact specifically with Site I sequences.
authors.affiliation: Department of Cell Biology, University of Massachusetts Medical School, Worcester 01655, USA.
authors.forename: K L
authors.initials: KL
authors.lastname: Wright
authors.forename: M J
authors.initials: MJ
authors.lastname: Birnbaum
authors.forename: A J
authors.initials: AJ
authors.lastname: van Wijnen
authors.forename: G S
authors.initials: GS
authors.lastname: Stein
authors.forename: J L
authors.initials: JL
authors.lastname: Stein
mesh_terms: D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D003432:Cross-Linking Reagents; D004247:DNA / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006657:Histones / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism / Q000528:radiation effects; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D014466:Ultraviolet Rays
chemical_list: D003432:Cross-Linking Reagents; D006657:Histones; D009687:Nuclear Proteins; D004247:DNA