Characterisation of functional domains in fission yeast Ams2 that are required for core histone gene transcription

Takayama, Yuko; Shirai, Masaki; Masuda, Fumie

doi:10.1038/srep38111

Cited by 6 publications

(10 citation statements)

References 29 publications

(68 reference statements)

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“…To determine whether reduced levels of Ams2 is important for other mitotic defects caused by H2Bub1 loss, we introduced plasmids expressing HA-tagged Ams2 from the nmt1 + promoter (which drives constitutive expression in media lacking thiamine) into either wild-type or htb1-K119R cells (Takayama et al 2016). Immunoblotting confirmed expres-sion of HA-Ams2 in both strains, although we consistently observed lower levels of expression in htb1-K119R cells (Figure S6A).…”

Section: Interplay Between H2bub1 and Cdk9 Regulates Ams2 + Antisensementioning

confidence: 80%

“…S. pombe strains were cultured in YE (0.5% yeast extract, 3% dextrose) or in EMM [Edinburgh minimal media; (Moreno et al 1991)] supplemented with adenine, leucine, uracil, and histidine (0.25 g/liter each). Strains harboring plasmids expressing ams2 + (Takayama et al 2016) were cultured in supplemented EMM lacking leucine and thiamine. Transformation of plasmids was carried out using a lithium acetate method (Bähler et al 1998).…”

Section: Methodsmentioning

confidence: 99%

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Histone H2B Ubiquitylation Regulates Histone Gene Expression by Suppressing Antisense Transcription in Fission Yeast

et al. 2019

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Histone H2B monoubiquitylation (H2Bub1) is tightly linked to RNA polymerase II transcription elongation, and is also directly implicated in DNA replication and repair. Loss of H2Bub1 is associated with defects in cell cycle progression, but how these are related to its various functions, and the underlying mechanisms involved, is not understood. Here we describe a role for H2Bub1 in the regulation of replication-dependent histone genes in the fission yeast Schizosaccharomyces pombe. H2Bub1 activates histone genes indirectly by suppressing antisense transcription of ams2 +-a gene encoding a GATA-type transcription factor that activates histone genes and is required for assembly of centromeric chromatin. Mutants lacking the ubiquitylation site in H2B or the H2B-specific E3 ubiquitin ligase Brl2 had elevated levels of ams2 + antisense transcripts and reduced Ams2 protein levels. These defects were reversed upon inhibition of Cdk9-an ortholog of the kinase component of positive transcription elongation factor b (P-TEFb)-indicating that they likely resulted from aberrant transcription elongation. Reduced Cdk9 activity also partially rescued chromosome segregation phenotypes of H2Bub1 mutants. In a genome-wide analysis, loss of H2Bub1 led to increased antisense transcripts at over 500 proteincoding genes in H2Bub1 mutants; for a subset of these, including several genes involved in chromosome segregation and chromatin assembly, antisense derepression was Cdk9-dependent. Our results highlight antisense suppression as a key feature of cell cycledependent gene regulation by H2Bub1, and suggest that aberrant transcription elongation may underlie the effects of H2Bub1 loss on cell cycle progression.

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Section: Interplay Between H2bub1 and Cdk9 Regulates Ams2 + Antisensementioning

confidence: 80%

Section: Methodsmentioning

confidence: 99%

Histone H2B Ubiquitylation Regulates Histone Gene Expression by Suppressing Antisense Transcription in Fission Yeast

et al. 2019

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“…SsAMS2 is homologous to protein AMS2, which has been demonstrated to be involved in activation the core histone genes expression at S phase of cell cycle in S. pombe (Takayama et al, 2016). In ams2 -deficient cell, the division cycle is longer than that in wild-type cells (Takayama and Takahashi, 2007).…”

Section: Resultsmentioning

confidence: 99%

“…GATA type zinc finger transcription factor (TF) was widely found in eukaryotes, plays critical roles in regulating physiological metabolism and controlling multicellular development (Lowry and Atchley, 2000; Liu et al, 2018). The ams2 gene, a m ulticopy s uppressor for the cpn1 mutant in fission yeast (Chen et al, 2003), encodes a GATA factor, binds to the histone gene promoter regions and takes part in transcriptional activation of the histone genes (Takayama et al, 2016). In Schizosaccharomyces pombe , AMS2, assembled in promoter regions of putative target genes and enriched in central centromeres, is critical for chromosome segregation, possibly via regulating localization of SpCENP-A ( cpn1 ) which is the centromere-specific histone H3 variant (Takayama et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Ssams2, a Gene Encoding GATA Transcription Factor, Is Required for Appressoria Formation and Chromosome Segregation in Sclerotinia sclerotiorum

et al. 2018

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AMS2, a multicopy suppressor for the cpn1 (SpCENP-A) mutant, functions to specifically regulate histone genes transcription and chromosome segregation. As a cell-cycle-regulated GATA transcription factor in eukaryotic organisms, little research has been done on the role of AMS2 protein in pathogenic fungi. In Sclerotinia sclerotiorum, Ssams2 (SS1G_03252) encodes a protein which has been predicted to contain GATA-box domain. Here, Ssams2-silenced strains with significantly reduced Ssams2 gene expression levels exhibited defect in hyphal growth, hyphal branching patterns, compound appressoria differentiation and the oxalic acid production compared to the wild-type (WT) strain. By common bean leaves infection assays, we identified the role of Ssams2 in full virulence. Furthermore, the numbers of cell nucleus in the same length of mycelium in Ssams2-silenced transformants were significantly less than that in the WT strain. The expression levels of histone genes and cell cycle genes in transformants were down-regulated significantly in the RNAi strains. Taken together, our work suggests that the TF SsAMS2 is required for growth, appressoria formation, virulence, and chromosome segregation in S. sclerotiorum.

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“…The ChIP assay was performed as described in [21]. The cells used in Sp525 cells containing pGP8110-Cnp1 or pGP4110-CENP-A L.s.…”

Section: Chromatin Immunoprecipitation (Chip) Assaymentioning

confidence: 99%

Identification of Genes Encoding CENP-A and Heterochromatin Protein 1 of Lipomyces starkeyi and Functional Analysis Using Schizosaccharomyces pombe

Takayama

2020

Genes

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Centromeres function as a platform for the assembly of multiple kinetochore proteins and are essential for chromosome segregation. An active centromere is characterized by the presence of a centromere-specific histone H3 variant, CENP-A. Faithful centromeric localization of CENP-A is supported by heterochromatin in almost all eukaryotes; however, heterochromatin proteins have been lost in most Saccharomycotina. Here, identification of CENP-A (CENP-AL.s.) and heterochromatin protein 1 (Lsw1) in a Saccharomycotina species, the oleaginous yeast Lipomyces starkeyi, is reported. To determine if these proteins are functional, the proteins in S. pombe, a species widely used to study centromeres, were ectopically expressed. CENP-AL.s. localizes to centromeres and can be replaced with S. pombe CENP-A, indicating that CENP-AL.s. is a functional centromere-specific protein. Lsw1 binds at heterochromatin regions, and chromatin binding is dependent on methylation of histone H3 at lysine 9. In other species, self-interaction of heterochromatin protein 1 is thought to cause folding of chromatin, triggering transcription repression and heterochromatin formation. Consistent with this, it was found that Lsw1 can self-interact. L. starkeyi chromatin contains the methylation of histone H3 at lysine 9. These results indicated that L. starkeyi has a primitive heterochromatin structure and is an attractive model for analysis of centromere heterochromatin evolution.

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Characterisation of functional domains in fission yeast Ams2 that are required for core histone gene transcription

Cited by 6 publications

References 29 publications

Histone H2B Ubiquitylation Regulates Histone Gene Expression by Suppressing Antisense Transcription in Fission Yeast

Histone H2B Ubiquitylation Regulates Histone Gene Expression by Suppressing Antisense Transcription in Fission Yeast

Ssams2, a Gene Encoding GATA Transcription Factor, Is Required for Appressoria Formation and Chromosome Segregation in Sclerotinia sclerotiorum

Identification of Genes Encoding CENP-A and Heterochromatin Protein 1 of Lipomyces starkeyi and Functional Analysis Using Schizosaccharomyces pombe

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