Asymmetric binding of histone H1 stabilizes MMTV nucleosomes and the interaction of progesterone receptor with the exposed HRE.
Packaging of mouse mammary tumor virus (MMTV) promoter sequences in nucleosomes modulates access of DNA binding proteins and influences the interaction among DNA bound transcription factors. Here we analyze the binding of histone H1 to MMTV mononucleosomes assembled with recombinant histones and study its influence on nucleosome structure and stability as well as on progesterone receptor (PR) binding to the hormone responsive elements (HREs). The MMTV nucleosomes can be separated into three main populations, two of which exhibited precise translational positioning. Histone H1 bound preferentially to the 5' distal nucleosomal DNA protecting additional 27-28 nt from digestion by micrococcal nuclease. Binding of histone H1 was unaffected by prior crosslinking of protein and DNA in nucleosomes with formaldehyde. Neither the translational nor the rotational nucleosome positioning was altered by histone H1 binding, but the nucleosomes were stabilized as judged by the kinetics of nuclease cleavage. Unexpectedly, binding of recombinant PR to the exposed distal HRE-I in nucleosomes was enhanced in the presence of histone H1, as demonstrated by band shift and footprinting experiments. This enhanced PR affinity may contribute to the reported positive effect of histone H1 on the hormonal activation of MMTV reporter genes.
Vicent
Guillermo P
GP
Institüt für Molekularbiologie und Tumorforschung (IMT), Philipps-Universität, Emil-Mannkoppf-Str. 2, D-35033, Marburg, Germany.
Meliá
María J
MJ
Beato
Miguel
M
D001665:Binding Sites; D003432:Cross-Linking Reagents / Q000737:chemistry; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics / Q000378:metabolism*; D009707:Nucleosomes / Q000737:chemistry / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011980:Receptors, Progesterone / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020218:Response Elements
D003432:Cross-Linking Reagents; D006657:Histones; D009707:Nucleosomes; D011980:Receptors, Progesterone; D011994:Recombinant Proteins