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Molecular and Cellular Biology, April 2006, p. 2965-2975, Vol. 26, No. 8
0270-7306/06/$08.00+0     doi:10.1128/MCB.26.8.2965-2975.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Genome-Wide Analysis of mRNAs Regulated by Drosha and Argonaute Proteins in Drosophila melanogaster{dagger}

Jan Rehwinkel,1 Pavel Natalin,1 Alexander Stark,1,{ddagger} Julius Brennecke,1 Stephen M. Cohen,1 and Elisa Izaurralde1,2*

EMBL, Meyerhofstrasse 1, D-69117 Heidelberg, Germany,1 MPI for Developmental Biology, Spemannstrasse 35, D-72076 Tübingen, Germany2

Received 13 October 2005/ Returned for modification 22 November 2005/ Accepted 3 February 2006

RNA silencing pathways are conserved gene regulation mechanisms that elicit decay and/or translational repression of mRNAs complementary to short interfering RNAs and microRNAs (miRNAs). The fraction of the transcriptome regulated by these pathways is not known, but it is thought that each miRNA may have hundreds of targets. To identify transcripts regulated by silencing pathways at the genomic level, we examined mRNA expression profiles in Drosophila melanogaster cells depleted of four Argonaute paralogs (i.e., AGO1, AGO2, PIWI, or Aubergine) that play essential roles in RNA silencing. We also profiled cells depleted of the miRNA-processing enzyme Drosha. The results reveal that transcripts differentially expressed in Drosha-depleted cells have highly correlated expression in the AGO1 knockdown and are significantly enriched in predicted and validated miRNA targets. The levels of a subset of miRNA targets are also regulated by AGO2. Moreover, AGO1 and AGO2 silence the expression of a common set of mobile genetic elements. Together, these results indicate that the functional overlap between AGO1 and AGO2 in Drosophila is more important than previously thought.

* Corresponding author. Mailing address: EMBL, Meyerhofstrasse 1, D-69117 Heidelberg, Germany. Phone: 49 6221 387 103. Fax: 49 6221 387 306. E-mail: izaurralde{at}embl-heidelberg.de.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

{ddagger} Present address: Broad Institute of MIT and Harvard, Cambridge, MA 02141.

Molecular and Cellular Biology, April 2006, p. 2965-2975, Vol. 26, No. 8
0270-7306/06/$08.00+0     doi:10.1128/MCB.26.8.2965-2975.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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