GSBS Student Publications

Title

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

UMMS Affiliation

Department of Biochemistry and Molecular Pharmacology

Date

9-17-2004

Document Type

Article

Medical Subject Headings

Base Sequence; DNA Primers; Flow Cytometry; Gene Silencing; Green Fluorescent Proteins; Luminescent Proteins; Schizosaccharomyces; Schizosaccharomyces pombe Proteins; Transcription, Genetic

Disciplines

Life Sciences | Medicine and Health Sciences

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.

Rights and Permissions

Citation: Genes Dev. 2004 Oct 1;18(19):2359-67. Epub 2004 Sep 15. Link to article on publisher's site

DOI of Published Version

10.1101/gad.1218004

Related Resources

Link to Article in PubMed

Journal Title

Genes and development

PubMed ID

15371329