Two DNA-binding sites on the globular domain of histone H5 are required for binding to both bulk and 5 S reconstituted nucleosomes
- PMID: 11071807
- DOI: 10.1006/jmbi.2000.4205
Two DNA-binding sites on the globular domain of histone H5 are required for binding to both bulk and 5 S reconstituted nucleosomes
Abstract
We have previously shown the existence of two DNA-binding sites on the globular domain of H5 (termed GH5), both of which are required for nucleosome organisation, as judged by the protection of a 166 bp chromatosome intermediate during micrococcal nuclease digestion of chromatin. This supports a model in which GH5 contacts two duplexes on the nucleosome. However, studies of a nucleosome assembled on the 5 S rRNA gene have argued against the requirement for two DNA-binding sites for chromatosome protection, which has implications for the role of linker histones. We have used this proposed difference in the requirement for a second site on the globular domain in the two models as a means of investigating whether bulk and reconstituted 5 S nucleosomes are indeed fundamentally different. GH5 protects a 166 bp chromatosome in both "bulk" and 5 S systems, and in both cases protection is abolished when all four basic residues in site II are replaced by alanine. Binding to four-way DNA junctions, which present a pair of juxtaposed duplexes, is also abolished. Single mutations of the basic residues did not abolish chromatosome protection in either system, or binding to four-way junctions, suggesting that the residues function as a cluster. Both bulk and 5 S nucleosomes thus require a functional second DNA-binding site on GH5 in order to bind properly to the nucleosome. This is likely to reflect a similar mode of binding in each case, in which two DNA duplexes are contacted in the nucleosome. There is no indication from these experiments that linker histones bind fundamentally differently to 5 S and bulk nucleosomes.
Copyright 2000 Academic Press.
Similar articles
-
Site-directed cleavage of DNA by a linker histone--Fe(II) EDTA conjugate: localization of a globular domain binding site within a nucleosome.Biochemistry. 1996 Sep 17;35(37):11931-7. doi: 10.1021/bi961590+. Biochemistry. 1996. PMID: 8810896
-
A K52Q substitution in the globular domain of histone H1t modulates its nucleosome binding properties.FEBS Lett. 2006 Oct 30;580(25):5999-6006. doi: 10.1016/j.febslet.2006.09.061. Epub 2006 Oct 5. FEBS Lett. 2006. PMID: 17052712
-
Crystal structure of globular domain of histone H5 and its implications for nucleosome binding.Nature. 1993 Mar 18;362(6417):219-23. doi: 10.1038/362219a0. Nature. 1993. PMID: 8384699
-
Xenopus transcription factor IIIA and the 5S nucleosome: development of a useful in vitro system.Biochem Cell Biol. 2003 Jun;81(3):177-84. doi: 10.1139/o03-043. Biochem Cell Biol. 2003. PMID: 12897852 Review.
-
DNA recognition and nucleosome organization.Biopolymers. 1997;44(4):423-33. doi: 10.1002/(SICI)1097-0282(1997)44:4<423::AID-BIP6>3.0.CO;2-M. Biopolymers. 1997. PMID: 9782778 Review.
Cited by
-
DNA sequence-dependent positioning of the linker histone in a nucleosome: A single-pair FRET study.Biophys J. 2021 Sep 7;120(17):3747-3763. doi: 10.1016/j.bpj.2021.07.012. Epub 2021 Jul 20. Biophys J. 2021. PMID: 34293303 Free PMC article.
-
Raw nuclear magnetic resonance data of human linker histone H1x, lacking the C-terminal domain (NGH1x), and trajectory data of nanosecond molecular dynamics simulations of GH1x- and NGH1x-chromatosomes.Data Brief. 2020 Jun 16;31:105865. doi: 10.1016/j.dib.2020.105865. eCollection 2020 Aug. Data Brief. 2020. PMID: 32642505 Free PMC article.
-
Histone H1 acetylation at lysine 85 regulates chromatin condensation and genome stability upon DNA damage.Nucleic Acids Res. 2018 Sep 6;46(15):7716-7730. doi: 10.1093/nar/gky568. Nucleic Acids Res. 2018. PMID: 29982688 Free PMC article.
-
Histone H5 is a potent Antimicrobial Agent and a template for novel Antimicrobial Peptides.Sci Rep. 2018 Feb 5;8(1):2411. doi: 10.1038/s41598-018-20912-1. Sci Rep. 2018. PMID: 29402952 Free PMC article.
-
Emerging roles of linker histones in regulating chromatin structure and function.Nat Rev Mol Cell Biol. 2018 Mar;19(3):192-206. doi: 10.1038/nrm.2017.94. Epub 2017 Oct 11. Nat Rev Mol Cell Biol. 2018. PMID: 29018282 Free PMC article. Review.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources