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PMID: 15371329 Published · ppublish English Journal Article

A single Argonaute protein mediates both transcriptional and posttranscriptional silencing in Schizosaccharomyces pombe.

Genes & development ·Vol. 18 ·No. 19 ·2004-10-01 ·Pages 2359-67

Sigova A, Rhind N, Zamore PD

Abstract

The Schizosaccharomyces pombe genome encodes only one of each of the three major classes of proteins implicated in RNA silencing: Dicer (Dcr1), RNA-dependent RNA polymerase (RdRP; Rdp1), and Argonaute (Ago1). These three proteins are required for silencing at centromeres and for the initiation of transcriptionally silent heterochromatin at the mating-type locus. Here, we show that the introduction of a double-stranded RNA (dsRNA) hairpin corresponding to a green fluorescent protein (GFP) transgene triggers classical RNA interference (RNAi) in S. pombe. That is, GFP silencing triggered by dsRNA reflects a change in the steady-state concentration of GFP mRNA, but not in the rate of GFP transcription. RNAi in S. pombe requires dcr1, rdp1, and ago1, but does not require chp1, tas3, or swi6, genes required for transcriptional silencing. Thus, the RNAi machinery in S. pombe can direct both transcriptional and posttranscriptional silencing using a single Dicer, RdRP, and Argonaute protein. Our findings suggest that these three proteins fulfill a common biochemical function in distinct siRNA-directed silencing pathways.

MeSH Terms
Base Sequence DNA Primers Flow Cytometry Gene Silencing/physiology Green Fluorescent Proteins Luminescent Proteins/genetics Schizosaccharomyces/genetics Schizosaccharomyces pombe Proteins/physiology Transcription, Genetic/physiology
Chemicals
DNA Primers Luminescent Proteins Schizosaccharomyces pombe Proteins Green Fluorescent Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Sigova Alla
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
Rhind Nicholas
Zamore Phillip D
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.1218004
Published
2004-10-01
Epub
2004-00-15
Pages
2359-67
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC522986
Subset
IM
External Links
PubMed source DOI: 10.1101/gad.1218004 PMC Full Text View journal details
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