title: Mobilization of hyperacetylated mononucleosomes by purified yeast ISW2 in vitro.
abstract: Catalytic activity of ISWI chromatin remodelers, which regulate nucleosome positioning on the DNA, depends on interactions of the putative acidic patch in ISWI helicase domain with the N-termini of nucleosomal H4--such, that removal of H4 termini abolishes ISWI remodeling. Acetylation of H4 termini is also known to disrupt H4 interactions with acidic protein surfaces, and thus, histone acetylation could potentially impede ISWI functions. Since active chromatin in vivo is hyperacetylated, it is important to clarify if ISWI activities can function on the in vivo hyperacetylated nucleosomes. We evaluated if purified yeast ISW2 can act on mononucleosomes in which all four core histones are highly acetylated. Mononucleosomes were assembled using purified histones from mammalian CV1 cells grown in the presence of deacetylase inhibitor Trichostatin A (TSA). The CV1 cell line is characterized by fast kinetic of accumulation of highly acetylated histone isoforms in response to TSA treatment. However, such 'native' histone hyperacetylation had no apparent effects on the nucleosome remodeling propensities, suggesting that histone hyperacetylation does not necessarily block ISWI functions and that ISWI enzymes can function on active chromatin as well.
authors.affiliation: Koltzov Institute of Developmental Biology of Russian Academy of Sciences, Moscow, 119334, Russia. Electronic address: wkrajewski@hotmail.com.
authors.forename: Wladyslaw A
authors.initials: WA
authors.lastname: Krajewski
mesh_terms: D000107:Acetylation; D000251:Adenosine Triphosphatases / Q000494:pharmacology*; D002384:Catalysis; D042002:Chromatin Assembly and Disassembly / Q000187:drug effects / Q000502:physiology*; D006657:Histones / Q000378:metabolism*; D009707:Nucleosomes / Q000187:drug effects* / Q000378:metabolism*; D014157:Transcription Factors / Q000494:pharmacology*
chemical_list: D006657:Histones; C086468:ISWI protein; D009707:Nucleosomes; D014157:Transcription Factors; D000251:Adenosine Triphosphatases
keywords: Active chromatin; Histone acetylation; ISWI remodeling; Nucleosomes; Transcription