Open AccessIdentification of Histone Mutants That Are Defective for Transcription-Coupled Nucleosome Occupancy
Author(s) -
Sarah J. Hainer,
Joseph A. Martens
Publication year - 2011
Publication title -
molecular and cellular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.14
H-Index - 327
eISSN - 1067-8824
pISSN - 0270-7306
DOI - 10.1128/mcb.05195-11
Subject(s) - biology , histone h3 , nucleosome , histone methylation , histone code , histone octamer , histone h2a , histone , genetics , histone h4 , chromatin immunoprecipitation , histone methyltransferase , transcription (linguistics) , promoter , microbiology and biotechnology , gene , gene expression , dna methylation , linguistics , philosophy
Our previous studies ofSaccharomyces cerevisiae described a gene repression mechanism where the transcription of intergenic noncoding DNA (ncDNA) (SRG1 ) assembles nucleosomes across the promoter of the adjacentSER3 gene that interfere with the binding of transcription factors. To investigate the role of histones in this mechanism, we screened a comprehensive library of histone H3 and H4 mutants for those that derepressSER3 . We identified mutations altering eight histone residues (H3 residues V46, R49, V117, Q120, and K122 and H4 residues R36, I46, and S47) that strongly increaseSER3 expression without reducing the transcription of the intergenicSRG1 ncDNA. We detected reduced nucleosome occupancy acrossSRG1 in these mutants to degrees that correlate well with the level ofSER3 derepression. The histone chromatin immunoprecipitation experiments on several other genes suggest that the loss of nucleosomes in these mutants is specific to highly transcribed regions. Interestingly, two of these histone mutants, H3 R49A and H3 V46A, reduce Set2-dependent methylation of lysine 36 of histone H3 and allow transcription initiation from cryptic intragenic promoters. Taken together, our data identify a new class of histone mutants that is defective for transcription-dependent nucleosome occupancy.