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Interdependent roles for histone chaperones and a chromatin boundary regulator in histone gene repression

ABSTRACT: A two-colour cell array screen reveals interdependent roles for histone chaperones and a chromatin boundary regulator in histone gene repression. We describe a fluorescent reporter system that exploits the functional genomic tools available in budding yeast to systematically assess consequences of genetic perturbations on gene expression. We used our Reporter-Synthetic Genetic Array (R-SGA) method to screen for regulators of core histone gene expression. We discovered that the histone chaperone Rtt106 functions in a pathway with two other chaperones, Asf1 and the HIR complex, to create a repressive chromatin structure at core histone promoters. We found that activation of histone (HTA1) gene expression involves both relief of Rtt106-mediated repression by the activity of the histone acetyltransferase Rtt109 and restriction of Rtt106 to the promoter region by the bromodomain-containing protein Yta7. We propose that the maintenance of Asf1/HIR/Rtt106-mediated repressive chromatin domains is the primary mechanism of cell cycle regulation of histone promoters. Our data suggest that this pathway may represent a chromatin regulatory mechanism that is broadly used across the genome.

ORGANISM(S): Saccharomyces cerevisiae

PROVIDER: GSE16693 | GEO | 2009/08/14

SECONDARY ACCESSION(S): PRJNA117375

REPOSITORIES: GEO

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A link between Sas2-mediated H4 K16 acetylation, chromatin assembly in S-phase by CAF-I and Asf1, and nucleosome assembly by Spt6 during transcription

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Transcriptome profiling reveals a novel role for Trichostatin A in antagonizing histone chaperone Chz1 mediated telomere anti-silencing.

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Transcriptome profiling reveals a novel role for Trichostatin A in antagonizing histone chaperone Chz1 mediated telomere anti-silencing.

Project description:The histone chaperones play an important role in chromatin assembly and disassembly during replication and transcription. We assessed the global roles of histone chaperones in Saccharomyces cerevisiae. Microarray transcriptional analyses indicate that histone chaperones have their own specific target genes, and various histone chaperones have partially overlapping functions during transcriptional regulation. Histone deacetylase inhibitor TSA and histone chaperones Asf1, Vps75 and Rtt106 can function in parallel pathways to regulate transcription. Moreover, TSA can specifically antagonize histone chaperone Chz1-mediated telomere anti-silencing. This study demonstrates that a mutual cross-talk mechanism exists between histone chaperones and histone deacetylation in transcriptional regulation. All yeast strains used in this study are listed in the paper (Table S1). CHZ1, NAP1, ASF1, VPS75 and RTT106 genes were deleted individually by homologous recombination in BY4742 (WT) by using a previously described in (Wan, Y. et al. MCB, 2009) PCR-based procedure. The strains were cultured at 30°C in YPD (1% yeast extract, 2% peptone, 2% glucose) media. For TSA treatments, WT and deletion mutants were grown to mid-log phase (OD600=0.5), TSA (Sigma-Aldrich) was then added to the yeast cultures at a final concentration of 10 ?M and cells were cultured for additional hour.Total RNA was isolated by hot acid phenol extraction protocol as previously described in Wan, Y. et al. MCB, 2009. Microarray labeling and hybridization reactions were performed as previously described in Wan, Y. et al. MCB, 2009 and Wan, Y. et al. NAR, 2010. Two color microarrays, comparing RNA from the experimental conditions (deletion mutations grown in YPD, WT and deletion mutations grown in YPD with TSA treatment) to RNA from the control WT cells grown in glucose-containing medium (YPD), were performed using Agilent whole-genome S. cerevisiae arrays.

2011-06-28 | E-GEOD-30228 | biostudies-arrayexpress

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Cell cycle- and chaperone-mediated regulation of H3K56ac incorporation in yeast

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SWI/SNF and the histone chaperone Rtt106 drive expression of the Pleiotropic Drug Resistance network genes

Project description:Genome-wide binding of the histone chaperone Rtt106 was analysed by ChIP-seq. Rtt106 binding was analysed in WT and the mutant backgrounds lacking transcription factors Pdr1 and Pdr3, the HIR histone chaperone subunit Hir1, or the boundary protein Yta7. Two biological replicates had been submitted. mRNA profiles in WT and the mutants lacking Rtt106, Pdr1, Pdr3 or the SWI/SNF subunit Snf2 were analysed by RNA-seq. mRNA profiles in those yeast cells grown in rich media (YPD) and in the glucose-starved condition (YEP) and in response to ketoconazole (YEP+ket) were analysed. Three biological samples for each condition had been submitted.

2022-02-21 | E-MTAB-9787 | biostudies-arrayexpress

Histone Recycling by FACT and Spt6 during Transcription Prevents the Scrambling of Histone Modifications [ChIP-seq]

Project description:The genetic information encoded in DNA is framed by additional layers of information, referred to as the epigenome. Epigenetic marks such as DNA methylation, histone modifications and histone variants are concentrated on specific genomic sites as means to instruct, but also sometimes as a consequence of, gene expression. How this information is maintained, notably in the face of transcription, is not understood. Here we show that the histone chaperones FACT and Spt6 are required for maintaining proper localization of several histone modifications including H3K4me1,2,3, H3K36me3, H3K79me3, H3K14ac, H3K18ac and H2Bub in Saccharomyces cerevisiae. In the absence of functional FACT or Spt6, transcription generates massive nucleosome loss which is partially compensated by increased histone assembly by histone chaperones such as Asf1 and HIR. Because re-incorporation of histones by these histone chaperones in not coupled to transcription, the modified histones are randomly incorporated, leading to scrambling of the epigenetic information. Hence, our work highlights the importance of local nucleosome recycling by FACT and Spt6 during transcription in the maintenance of a proper epigenetic landscape.

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