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Mkt1 is required for RNAi-mediated silencing and establishment of heterochromatin in fission yeast [CRAC]

ABSTRACT: Mkt1 is required for RNAi-mediated silencing and establishment of heterochromatin in fission yeast [CRAC]

PROVIDER: PRJNA559880 | ENA |

REPOSITORIES: ENA

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Mkt1 is required for RNAi-mediated silencing and establishment of heterochromatin in fission yeast

Project description:Constitutive domains of repressive heterochromatin are maintained within the fission yeast genome through self-reinforcing mechanisms involving histone methylation and small RNAs. Non-coding RNAs generated from heterochromatic regions are processed into small RNAs by the RNA interference pathway, and are subject to silencing through both transcriptional and post-transcriptional mechanisms. While the pathways involved in maintenance of the repressive heterochromatin state are reasonably well understood, less is known about the requirements for its establishment. Here we describe a novel role for the post-transcriptional regulatory factor Mkt1 in establishment of heterochromatin at pericentromeres in fission yeast. Loss of Mkt1 does not affect maintenance of existing heterochromatin, but does affect its recovery following depletion, as well as de novo establishment of heterochromatin on a mini-chromosome. Pathway dissection revealed that Mkt1 is required for RNAi-mediated post-transcriptional silencing, downstream of small RNA production. Mkt1 physically associates with pericentromeric transcripts, and is additionally required for maintenance of silencing and heterochromatin at centromeres when transcriptional silencing is impaired. Our findings provide new insight into the mechanism of RNAi-mediated post-transcriptional silencing in fission yeast, and unveil an important role for post-transcriptional silencing in establishment of heterochromatin that is dispensable when full transcriptional silencing is imposed.

2019-11-28 | PXD015484 | Pride

Mkt1 is required for RNAi-mediated silencing and establishment of heterochromatin in fission yeast [CRAC]

Project description:CRAC analysis was performed to identify transcripts associated with the Mkt1 protein in Schizosaccharomyces pombe

2019-11-28 | GSE135735 | GEO

Mkt1 is required for RNAi-mediated silencing and establishment of heterochromatin in fission yeast [RNA-seq]

Project description:Mkt1 is required for RNAi-mediated silencing and establishment of heterochromatin in fission yeast [RNA-seq]

| PRJNA558143 | ENA

Mkt1 is required for RNAi-mediated silencing and establishment of heterochromatin in fission yeast.

Project description: Not available

| S-EPMC7026591 | biostudies-literature

Genome wide map of heterochromatin state in fission yeast Schizosaccharomyces pombe

Project description:Genome wide map of heterochromatin state in fission yeast Schizosaccharomyces pombe via 4 different strains Examination of a single histone modification in 4 different fission yeast strains

2013-07-20 | E-GEOD-42848 | biostudies-arrayexpress

Nuclear retention of fission yeast dicer is a prerequisite for RNAi-mediated heterochromatin assembly

Project description:RNAseIII ribonucleases act at the heart of RNA silencing pathways by processing precursor RNAs into mature microRNAs and siRNAs. In the fission yeast Schizosaccharomyces pombe, siRNAs are generated by the RNAseIII enzyme Dcr1 and are required for heterochromatin formation. In this study, we have analyzed the subcellular localization of Dcr1 and found that it accumulates in the nucleus and is enriched at the nuclear periphery. Nuclear accumulation of Dcr1 depends on a short motif which constrains nuclear export promoted by the double-stranded RNA binding domain of Dcr1. Absence of this motif renders Dcr1 mainly cytoplasmic and is accompanied by remarkable changes in gene expression and failure to assemble heterochromatin. Our findings suggest that dicer proteins are shuttling proteins and that the steady-state subcellular levels can be shifted towards either compartment. This has implications for the mechanism of RNAi-mediated heterochromatin assembly and the spatial organization of RNA silencing pathways in general. Small RNA libraries from total RNA isolations of wild-type, dcr1Delta and dcr1DeltaC33 cells and subjected to high-throughput sequencing.

2010-02-16 | E-GEOD-18582 | biostudies-arrayexpress

Nuclear retention of fission yeast dicer is a prerequisite for RNAi-mediated heterochromatin assembly

2010-02-16 | GSE18582 | GEO

Nuclear retention of fission yeast dicer is a prerequisite for RNAi-mediated heterochromatin assembly

2010-02-16 | GSE18440 | GEO

Genome wide map of heterochromatin state in fission yeast Schizosaccharomyces pombe

Project description:Genome wide map of heterochromatin state in fission yeast Schizosaccharomyces pombe via 4 different strains

2013-07-20 | GSE42848 | GEO

Mediator directs co-transcriptional heterochromatin assembly by RNAi-dependent and -independent pathways.

Project description:Background: Heterochromatin at the pericentromeric repeats in fission yeast is assembled and spread by an RNAi-dependent mechanism, which is coupled to the transcription of non-coding RNA from the repeats by RNA polymerase II. In addition, Rrp6, a component of the nuclear exosome, also contributes to heterochromatin assembly and is coupled to non-coding RNA transcription. The multi-subunit complex Mediator, which directs initiation of RNA polymerase II-dependent transcription, has recently been suggested to function after initiation in processes such as elongation of transcription and splicing. However, the role of Mediator in the regulation of chromatin structure is not well understood. Results: We investigated the role of Mediator in pericentromeric heterochromatin formation and found that deletion of specific subunits of the head domain of Mediator compromised heterochromatin structure. The Mediator head domain was required for Rrp6-dependent heterochromatin nucleation at the pericentromere and for RNAi-dependent spreading of heterochromatin into the neighboring region. In the latter process, Mediator appeared to contribute to efficient processing of siRNA from transcribed non-coding RNA, which was required for efficient spreading of heterochromatin. Furthermore, the head domain directed efficient transcription in heterochromatin. Conclusions: These results reveal a pivotal role for Mediator in multiple steps of transcription-coupled formation of pericentromeric heterochromatin. This observation further extends the role of Mediator to co-transcriptional chromatin regulation. Gene expression profile at exponentially-growing phase.in the fission yeast deletion mutants of pmc6, med20 and dcr1.

2013-08-31 | E-GEOD-43543 | biostudies-arrayexpress

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