title: Characterization of nucleosome core particles containing histone proteins made in bacteria.
abstract: The four core histone proteins, H2A, H2B, H3, and H4 of Xenopus laevis have been individually expressed in milligram quantities in Escherichia coli. The full-length proteins and the "trypsin-resistant" globular domains were purified under denaturing conditions and folded into histone octamers. Both intact and truncated recombinant octamers, as well as chicken erythrocyte octamer, were assembled into nucleosome core particles using a 146 bp defined-sequence DNA fragment from a 5 S RNA gene. The three types of core particles were characterized and compared by gel electrophoresis, DNase I cleavage, and tyrosine fluorescence emission during stepwise dissociation with increasing ionic strength. Nucleosome core particles containing native and mutant histones made in bacteria have facilitated its X-ray structure determination at 2.8 A resolution.
authors.affiliation: Institut für Molekularbiologie und Biophysik, ETH-Hönggerberg, Zürich, CH-8093, Switzerland.
authors.forename: K
authors.initials: K
authors.lastname: Luger
authors.forename: T J
authors.initials: TJ
authors.lastname: Rechsteiner
authors.forename: A J
authors.initials: AJ
authors.lastname: Flaus
authors.forename: M M
authors.initials: MM
authors.lastname: Waye
authors.forename: T J
authors.initials: TJ
authors.lastname: Richmond
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002645:Chickens; D003001:Cloning, Molecular; D004912:Erythrocytes / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D005813:Genes, Synthetic; D006657:Histones / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009707:Nucleosomes / Q000737:chemistry*; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding; D017510:Protein Folding; D012341:RNA, Ribosomal, 5S; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D014982:Xenopus laevis / Q000235:genetics
chemical_list: D006657:Histones; D009707:Nucleosomes; D012341:RNA, Ribosomal, 5S; D011994:Recombinant Proteins