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HIRA, a Conserved Histone Chaperone, Plays an Essential Role in Low-dose Stress Response via Transcriptional Stimulation in Fission Yeast

ABSTRACT: Cells that have been pre-exposed to mild stress (priming stress) acquire transient resistance to subsequent severe stress even under different combinations of stresses. This phenomenon is called cross-tolerance. Although it has been reported that cross-tolerance occurs in many organisms, the molecular basis is not clear yet. Here, we identified slm9+ as a responsible gene for the cross-tolerance in the fission yeast Schizosaccharomyces pombe. Slm9 is a homolog of mammalian HIRA histone chaperone. HIRA forms a conserved complex and gene disruption of other HIRA complex components, Hip1, Hip3, and Hip4, also yielded a cross-tolerance-defective phenotype, indicating that the fission yeast HIRA is involved in the cross-tolerance as a complex. We also revealed that Slm9 was recruited to the stress-responsive gene loci upon stress treatment in an Atf1-dependent manner. The expression of stress-responsive genes under stress conditions was compromised in HIRA disruptants. Consistent with this, Pol II recruitment and nucleosome eviction at these gene loci were impaired in slm9D cells. Furthermore, we found that the priming stress enhanced the expression of stress-responsive genes in wild-type cells that were exposed to the severe stress. These observations suggest that HIRA functions in stress response through transcriptional regulation. To determine whether fission yeast HIRA specifically regulates stress-responsive genes under stress condition, we performed genome-wide analysis by using Affymetrix GeneChip oligonucleotide microarrays.

ORGANISM(S): Saccharomyces cerevisiae Schizosaccharomyces pombe

PROVIDER: GSE35281 | GEO | 2012/07/22

SECONDARY ACCESSION(S): PRJNA151575

REPOSITORIES: GEO

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HIRA, a Conserved Histone Chaperone, Plays an Essential Role in Low-dose Stress Response via Transcriptional Stimulation in Fission Yeast

Project description:Cells that have been pre-exposed to mild stress (priming stress) acquire transient resistance to subsequent severe stress even under different combinations of stresses. This phenomenon is called cross-tolerance. Although it has been reported that cross-tolerance occurs in many organisms, the molecular basis is not clear yet. Here, we identified slm9+ as a responsible gene for the cross-tolerance in the fission yeast Schizosaccharomyces pombe. Slm9 is a homolog of mammalian HIRA histone chaperone. HIRA forms a conserved complex and gene disruption of other HIRA complex components, Hip1, Hip3, and Hip4, also yielded a cross-tolerance-defective phenotype, indicating that the fission yeast HIRA is involved in the cross-tolerance as a complex. We also revealed that Slm9 was recruited to the stress-responsive gene loci upon stress treatment in an Atf1-dependent manner. The expression of stress-responsive genes under stress conditions was compromised in HIRA disruptants. Consistent with this, Pol II recruitment and nucleosome eviction at these gene loci were impaired in slm9D cells. Furthermore, we found that the priming stress enhanced the expression of stress-responsive genes in wild-type cells that were exposed to the severe stress. These observations suggest that HIRA functions in stress response through transcriptional regulation. To determine whether fission yeast HIRA specifically regulates stress-responsive genes under stress condition, we performed genome-wide analysis by using Affymetrix GeneChip oligonucleotide microarrays. Fission yeast cells (WT, slm9D, hip1D) were grown in quadruplicate at 32Â°C to the logarithmic phase and an aliquot was collected as the unstressed control. The other three aliquots were exposed to 40Â°C for 1 h, 25 mM H2O2 for 1 h, or 40Â°C for 1 h followed by 25 mM H2O2 for 1 h, respectively. Total RNA was purified and all the 12 RNA samples were analyzed with GeneChip Yeast Genome 2.0 Array (Affymetrix).

2012-07-21 | E-GEOD-35281 | biostudies-arrayexpress

Transcription profiling of hip1::ura4+ and slm9::ura4+ S. pombe to investigate the role of the HIRA histone chaperone complex

Project description:The assembly of nucleosomes by histone chaperones is an important component of transcriptional regulation. Here we have assessed the global roles of the S. pombe HIRA histone chaperone complex. Microarray analysis indicates that inactivation of the HIRA complex results in increased expression of at least 4% of fission yeast genes. HIRA-regulated genes overlap with those which are normally repressed in vegetatively growing cells, such as targets of the Clr6 histone deacetylase and silenced genes located in subtelomeric regions. HIRA is also required for silencing of all 13 intact copies of the Tf2 long terminal repeat (LTR) retrotransposon. However, the role of HIRA is not restricted to bona fide promoters, because it also suppresses non-coding transcripts from solo LTR elements and spurious antisense transcripts from cryptic promoters associated with transcribed regions. Furthermore, the HIRA complex is essential in the absence of the quality control provided by nuclear exosome-mediated degradation of illegitimate transcripts. This suggests that HIRA restricts genomic accessibility, and, consistent with this, the chromosomes of cells lacking HIRA are more susceptible to genotoxic agents that cause double strand breaks. Thus the HIRA histone chaperone is required to maintain the protective functions of chromatin.

2009-07-22 | E-MTAB-130 | biostudies-arrayexpress

Expression data from Control, HIRA and CABIN1 knockdown cells

Project description:The mammalian HIRA/UBN1/ASF1a complex is a histone chaperone complex that is conserved from yeast (Saccharomyces cerevisiae) to humans. This complex preferentially deposits the histone variant H3.3 into chromatin in a DNA replication-independent manner and is implicated in diverse chromatin regu- latory events from gene activation to heterochromatinization. In yeast, the orthologous complex consists of three Hir proteins (Hir1p, Hir2p, and Hir3p), Hpc2p, and Asf1p. Yeast Hir3p has weak homology to CABIN1, a fourth member of the human complex, suggesting that Hir3p and CABIN1 may be orthologs. Here we show that HIRA and CABIN1 interact at ectopic and endogenous levels of expression in cells, and we isolate the quaternary HIRA/UBN1/CABIN1/ASF1a (HUCA) complex, assembled from recombinant proteins. Mutational analyses support the view that HIRA acts as a scaffold to bring together UBN1, ASF1a, and CABIN1 into a quaternary complex. We show that, like HIRA, UBN1, and ASF1a, CABIN1 is involved in heterochromatinization of the genome of senescent human cells. Moreover, in proliferating cells, HIRA and CABIN1 regulate overlapping sets of genes, and these genes are enriched in the histone variant H3.3. In sum, these data demonstrate that CABIN1 is a functional member of the human HUCA complex and so is the likely ortholog of yeast Hir3p. We used microarrays to detail the global programme of gene expression after knockdown of HIRA and CABIN1 in 3 replicates HeLa cells were nucleofacted with Dharmacon control siRNA and siRNA to HIRA and CABIN1 and RNA was isolated 72 hours after transfection.

2013-03-20 | E-GEOD-45295 | biostudies-arrayexpress

Transcription profiling of fission yeast meiosis and sporulation

Project description:Fission yeast cells undergo meiosis and sporulation under conditions of nutritional stress, most frequently nitrogen starvation. This is a complex developmental process, which results in the formation of four spores that are highly resistant to environmental stress. We have carried out time course experiments with diploid fission yeast cells undergoing meiosis and sporulation. In this experiment we have used wild type diploids, and induced meiosis by removal of nitrogen from the medium. As a reference we used a pool consisting of equal amount of RNA from all time points of the experiments. This experiment is part of a series, with accession numbers E-SNGR-2 to E-SNGR-7.

2002-08-05 | E-SNGR-3 | biostudies-arrayexpress

Expression data from Control, HIRA and CABIN1 knockdown cells

2013-03-20 | GSE45295 | GEO

HDAC mediated suppression of histone turnover promotes epigenetic stability of heterochromatin

Project description:Analysis of histone H3 turnover in wild-type and mutant fission yeast cells by using MNase-ChIP-chip Exponentially growing cultures of fission yeast cells expressing an ectopic copy of FLAG tagged histone H3 under the control of inv1 promoter were synchronized and expression of H3-FLAG was induced by changing the carbon source of the medium to Sucrose. Cells were crosslinked with 1% Formaldehyde and chromatin was fragmented into mononucleosomes by using micrococcal nuclease (MNase). Chromatin immunoprecipitated DNA recovered with anti-FLAG antibody from wild-type and mutant fission yeast cells and whole cell extracts DNA were amplified by random priming and respectively labeled with Cy5 and Cy3. Labeled DNA samples were mixed and hybridized onto 44k pombe Agilent oligo arrays. Data were processed using Agilent scanner and Feature Extraction software.

2013-04-21 | E-GEOD-45378 | biostudies-arrayexpress

Schizosaccharomyces pombe

Project description:Expression data from fission yeast Schizosaccharomyces pombe under nitrosative stress

| PRJNA219319 | ENA

Transcription profiling of fission yeast meiosis and sporulation

Project description:Transcriptional profile of fission yeast meiosis and sporulation. Fission yeast cells undergo meiosis and sporulation under conditions of nutritional stress, most frequently nitrogen starvation. This is a complex developmental process, which results in the formation of four spores that are highly resistant to environmental stress. We have carried out time courses of diploid fission yeast cells undergoing meiosis and sporulation. To achieve the best possible synchrony we have used thermosensitive mutants in the meiotic inhibitor pat1. We first pre-synchronized the cells in G1 by removal of nitrogen at the permissive temperature for pat1, and then induced meiosis by inactivating pat1 with a temperature shift. pat1-induced meiosis is a standard technique, resulting in a highly synchronous meiosis that is very similar to the normal process. The use of pat1 mutants and a temperature-shift may cause some artifacts, so these results should be viewed together with temperature shift controls and especially with the results of the time course done with wild type diploids, available under accession numbers E-SNGR-3 to E-SNGR-7. In all cases we used as reference a pool consisting of equal amounts of RNA from all time points of the experiment. This experiment describes the first biological replicate for pat1 using array design A-SNGR-2. Note that there are three other biological replicates, which use array design A-SNGR-1 in experiment E-SNGR-2.

2002-08-05 | E-SNGR-7 | biostudies-arrayexpress

A global non coding RNA system modulates fission yeast protein levels in response to stress

Project description:Mitogen Activated Protein Kinase (MAPK) signaling cascades transduce information arising from events external to the cell, such as environmental stresses, to a variety of downstream effectors and transcription factors. The fission yeast stress activated MAP kinase (SAPK) pathway is conserved with the p38 and JNK pathways in humans, and comprises the MAPKKKs Win1, Wis4, the MAPKK Wis1, and the MAPK, Sty1. Sty1 and its main downstream effector Atf1 regulate a large set of core environmental stress response genes. The fission yeast genome encodes three other ATF proteins: Atf21, Atf31 and Pcr1. Among these, atf21 is specifically induced under conditions of high osmolarity. We have therefore instigated a programme to investigate the role played by non coding RNAs (ncRNAs) in response to osmotic stress challenge in wild type and atf21Δ cells. By integrating global proteomics and RNA sequencing data, we identified a systematic program in which elevated antisense RNAs arising both from ncRNAs and from 3'-overlapping convergent gene-pairs is directly associated with substantial reductions in protein levels throughout the fission yeast genome. We also found an extensive array of ncRNAs with trans associations that have the potential to influence different biological processes and stress responses in fission yeast, suggesting ncRNAs comprise additional components of the SAPK regulatory system.

2014-05-19 | GSE56761 | GEO

Transcription profiling of S. pombe to investigate genetic control of cellular quiescence

Project description:Transition from proliferation to quiescence brings about extensive changes in cellular behavior and structure. However, genes critical for establishing and/or for maintaining quiescence are largely unknown. The fission yeast S. pombe is found as an excellent model for studying this problem, because it becomes quiescent under nitrogen starvation. Here we characterize 610 temperature-sensitive (ts) mutants, and identify 33 genes required for entry into and the maintenance of quiescence. These genes cover a broad range of cellular functions in the cytoplasm, membrane and the nucleus, encoding proteins for stress-responsive and cell cycle kinase signaling pathway, actin-bound and osmo-controlling endosome formation, RNA transcription, splicing and ribosome biogenesis, chromatin silencing, biosynthesis of lipid and ATP, cell wall and membrane morphogenesis, protein trafficking and vesicle fusion. We specifically highlight Fcp1, CTD phosphatase of RNA polymerase II, which differentially affects transcription of genes involved in quiescence and proliferation. We propose that the transcriptional role of Fcp1 is central to differentiate quiescence from proliferation. Experiment Overall Design: Gene expression profile under nitrogen rich or starved condition at 26ÂºC or 37ÂºC in WT (L972) or fcp1-452 ts mutant strain of fission yeast.

2009-05-11 | E-GEOD-14319 | biostudies-arrayexpress

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