title: The site-specific installation of methyl-lysine analogs into recombinant histones.
abstract: Histone lysine residues can be mono-, di-, or trimethylated. These posttranslational modifications regulate the affinity of effector proteins and may also impact chromatin structure independent of their role as adaptors. In order to study histone lysine methylation, particularly in the context of chromatin, we have developed a chemical approach to install analogs of methyl lysine into recombinant proteins. This approach allows for the rapid generation of large quantities of histones in which the site and degree of methylation can be specified. We demonstrate that these methyl-lysine analogs (MLAs) are functionally similar to their natural counterparts. These methylated histones were used to examine the influence of specific lysine methylation on the binding of effecter proteins and the rates of nucleosome remodeling. This simple method of introducing site-specific and degree-specific methylation into recombinant histones provides a powerful tool to investigate the biochemical mechanisms by which lysine methylation influences chromatin structure and function.
authors.affiliation: Department of Chemistry, University of California, Berkeley, CA 94720, USA.
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Simon
authors.forename: Feixia
authors.initials: F
authors.lastname: Chu
authors.forename: Lisa R
authors.initials: LR
authors.lastname: Racki
authors.forename: Cecile C
authors.initials: CC
authors.lastname: de la Cruz
authors.forename: Alma L
authors.initials: AL
authors.lastname: Burlingame
authors.forename: Barbara
authors.initials: B
authors.lastname: Panning
authors.forename: Geeta J
authors.initials: GJ
authors.lastname: Narlikar
authors.forename: Kevan M
authors.initials: KM
authors.lastname: Shokat
mesh_terms: D000818:Animals; D003545:Cysteine / Q000737:chemistry; D006657:Histones / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008239:Lysine / Q000031:analogs & derivatives*; D008745:Methylation; D008958:Models, Molecular; D009707:Nucleosomes / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D029867:Xenopus Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D006657:Histones; D009707:Nucleosomes; D011994:Recombinant Proteins; D029867:Xenopus Proteins; D008239:Lysine; D003545:Cysteine