title: Interaction of chromatin with a histone H1 containing swapped N- and C-terminal domains.
abstract: Although the details of the structural involvement of histone H1 in the organization of the nucleosome are quite well understood, the sequential events involved in the recognition of its binding site are not as well known. We have used a recombinant human histone H1 (H1.1) in which the N- and C-terminal domains (NTD/CTD) have been swapped and we have reconstituted it on to a 208-bp nucleosome. We have shown that the swapped version of the protein is still able to bind to nucleosomes through its structurally folded wing helix domain (WHD); however, analytical ultracentrifuge analysis demonstrates its ability to properly fold the chromatin fibre is impaired. Furthermore, FRAP analysis shows that the highly dynamic binding association of histone H1 with the chromatin fibre is altered, with a severely decreased half time of residence. All of this suggests that proper binding of histone H1 to chromatin is determined by the simultaneous and synergistic binding of its WHD-CTD to the nucleosome.
authors.affiliation: Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia, Canada V8W 3P6.
authors.forename: Jordana B
authors.initials: JB
authors.lastname: Hutchinson
authors.affiliation: Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia, Canada V8W 3P6.
authors.forename: Manjinder S
authors.initials: MS
authors.lastname: Cheema
authors.affiliation: Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia, Canada V8W 3P6.
authors.forename: Jason
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Oncology, Faculty of Dentistry and Medicine, University of Alberta, Edmonton, Alberta, Canada T6G 1Z2.
authors.forename: Krystal
authors.initials: K
authors.lastname: Missiaen
authors.affiliation: Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia, Canada V8W 3P6.
authors.forename: Ron
authors.initials: R
authors.lastname: Finn
authors.affiliation: Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia, Canada V8W 3P6.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Gonzalez Romero
authors.affiliation: Northern Ontario School of Medicine, 955 Oliver Road, Thunder Bay, Ontario, Canada P7B 5E1.
authors.forename: John P H
authors.initials: JP
authors.lastname: Th'ng
authors.affiliation: Department of Oncology, Faculty of Dentistry and Medicine, University of Alberta, Edmonton, Alberta, Canada T6G 1Z2.
authors.forename: Michael
authors.initials: M
authors.lastname: Hendzel
authors.affiliation: Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia, Canada V8W 3P6 jausio@uvic.ca.
authors.forename: Juan
authors.initials: J
authors.lastname: Ausió
mesh_terms: D001665:Binding Sites; D002843:Chromatin / Q000378:metabolism* / Q000648:ultrastructure; D002942:Circular Dichroism; D006367:HeLa Cells; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D009707:Nucleosomes / Q000378:metabolism; D000072417:Protein Domains; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D002843:Chromatin; D006657:Histones; D009707:Nucleosomes; D011994:Recombinant Proteins
keywords: FRAP; analytical ultracentrifuge; chromatin; circular dichroism; histones