title: Site-directed mutagenesis studies on the binding of the globular domain of linker histone H5 to the nucleosome.
abstract: The globular domain of the linker histone H5 has been expressed in Escherichia coli. The purified peptide is functional as it permits chromatosome protection during micrococcal nuclease digestion of chromatin reconstituted with the peptide, indicating that it binds correctly at the dyad axis of the nucleosomal core particle. The globular domain residue lysine 64 is highly conserved within the linker histone family, and site-directed mutagenesis has been used to assess the importance of this residue in the binding of the globular domain of linker histone H5 to the nucleosome. Recombinant peptides mutated at lysine 64 are unable to elicit chromatosome protection to the same degree as the wild-type peptide, and since they appear to be fully folded, these observations confirm a major role for this residue in determining the effective interaction between the globular domain of histone H5 and the nucleosome.
authors.affiliation: Department of Biophysics, Cell and Molecular Biology, King's College, London, U.K.
authors.forename: R S
authors.initials: RS
authors.lastname: Buckle
authors.forename: J D
authors.initials: JD
authors.lastname: Maman
authors.forename: J
authors.initials: J
authors.lastname: Allan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002645:Chickens; D002843:Chromatin / Q000737:chemistry; D003001:Cloning, Molecular; D004247:DNA; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D006657:Histones / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009707:Nucleosomes / Q000378:metabolism*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014357:Trypsin
chemical_list: D002843:Chromatin; D006657:Histones; D009707:Nucleosomes; D011994:Recombinant Proteins; D004247:DNA; D014357:Trypsin