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Schizosaccharomyces pombe

ABSTRACT: Tailing and degradation of Argonaute-bound small RNAs protect the genome from uncontrolled RNAi

PROVIDER: PRJNA378525 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR5320760.fastq.gz	Fastqsanger.gz
	SRR5320770.fastq.gz	Fastqsanger.gz
	SRR5320780.fastq.gz	Fastqsanger.gz
	SRR5320761.fastq.gz	Fastqsanger.gz
	SRR5320771.fastq.gz	Fastqsanger.gz

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Tailing and degradation of Argonaute-bound small RNAs protect the genome from uncontrolled RNAi

Project description:RNAi is a conserved mechanism in which small RNAs induce silencing of complementary targets. How Argonaute-bound small RNAs are targeted for degradation is not well understood. We show here that the adenyl-transferase Cid14, a member of the TRAMP complex, and the uridyl-transferase Cid16 add non-templated nucleotides to Argonaute-bound small RNAs in fission yeast. The tailing of Argonaute-bound small RNAs in vitro recruits the 3’-5’ exonuclease Rrp6 to degrade small RNAs. Failure in degradation of Argonaute-bound small RNAs results in accumulation of “noise” small RNAs on Argonaute and targeting of diverse euchromatic genes by RNAi. To protect themselves from uncontrolled RNAi, cid14Δ cells exploit the RNAi machinery and silence genes essential for RNAi itself, which is required for their viability. Our data indicate that surveillance of Argonaute-bound small RNAs by Cid14/Cid16 and the exosome protects the genome from uncontrolled RNAi and reveal a rapid RNAi-based adaptation to stress conditions.

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RNAi mediates post-transcriptional repression of gene expression in fission yeast Schizosaccharomyces pombe [Affymetrix]

Project description:RNA interference (RNAi) is a gene silencing mechanism conserved from fungi to mammals. Small interfering RNAs are products and mediators of the RNAi pathway and act as specificity factors in recruiting effector complexes. The Schizosaccharomyces pombe genome encodes one of each of the core RNAi proteins, Dicer, Argonaute and RNA-dependent RNA polymerase (dcr1, ago1, rdp1). Even though the function of RNAi in heterochromatin assembly in S. pombe is established, its role in controlling gene expression is elusive. Here, we report the identification of small RNAs mapped anti-sense to protein coding genes in fission yeast. We demonstrate that these genes are up-regulated at the protein level in RNAi mutants, while their mRNA levels are not significantly changed. We show that the repression by RNAi is not a result of heterochromatin formation. Thus, we conclude that RNAi is involved in post-transcriptional gene silencing in S. pombe.

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TRAMP-mediated RNA surveillance prevents spurious entry of RNAs into the S. pombe siRNA pathway

Project description:In the fission yeast Schizosaccharomyces pombe, the RNAi machinery is required to generate small interfering RNAs (siRNAs) that mediate heterochromatic gene silencing. Efficient silencing also requires the TRAMP complex, which contains the non-canonical Cid14 polyA polymerase and targets aberrant RNAs for degradation. Here we use high-throughput sequencing to analyze Argonaute-associated small RNAs (sRNAs) in both the presence and absence of Cid14. Most sRNAs in fission yeast start with a 5’-uracil, largely because these are loaded most efficiently into Argonaute. In wild-type cells, most sRNAs match to repeated regions of the genome, whereas in cid14 cells, the sRNA profile changes to include major new classes of sRNAs originating from ribosomal RNAs and a tRNA. Thus, Cid14 prevents certain abundant RNAs from becoming substrates for the RNAi machinery, thereby freeing the RNAi machinery to act on its proper targets.

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Project description:Histone H2A.Z cooperates with RNAi and heterochromatin factors to suppress antisense RNAs

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2014-01-23 | E-GEOD-54194 | biostudies-arrayexpress

Schizosaccharomyces pombe