Comparative Analysis of Argonaute-Dependent Small RNA Pathways in Drosophila

Rui Zhou; Ikuko Hotta; Ahmet M. Denli; Pengyu Hong; Norbert Perrimon; Gregory J. Hannon

doi:10.1016/j.molcel.2008.10.018

Comparative Analysis of Argonaute-Dependent Small RNA Pathways in Drosophila

Rui Zhou, Ikuko Hotta, Ahmet M. Denli, Pengyu Hong, Norbert Perrimon, Gregory J. Hannon

Research output: Contribution to journal › Article › peer-review

92 Scopus citations

Abstract

The specificity of RNAi pathways is determined by several classes of small RNAs, which include siRNAs, piRNAs, endo-siRNAs, and microRNAs (miRNAs). These small RNAs are invariably incorporated into large Argonaute (Ago)-containing effector complexes known as RNA-induced silencing complexes (RISCs), which they guide to silencing targets. Both genetic and biochemical strategies have yielded conserved molecular components of small RNA biogenesis and effector machineries. However, given the complexity of these pathways, there are likely to be additional components and regulators that remain to be uncovered. We have undertaken a comparative and comprehensive RNAi screen to identify genes that impact three major Ago-dependent small RNA pathways that operate in Drosophila S2 cells. We identify subsets of candidates that act positively or negatively in siRNA, endo-siRNA, and miRNA pathways. Our studies indicate that many components are shared among all three Argonaute-dependent silencing pathways, though each is also impacted by discrete sets of genes.

Original language	English (US)
Pages (from-to)	592-599
Number of pages	8
Journal	Molecular cell
Volume	32
Issue number	4
DOIs	https://doi.org/10.1016/j.molcel.2008.10.018
State	Published - Nov 21 2008
Externally published	Yes

Keywords

PROTEINS
RNA

ASJC Scopus subject areas

Molecular Biology
Cell Biology

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10.1016/j.molcel.2008.10.018

Cite this

@article{c80f7eb0a0fa4521867293700ac9e380,

title = "Comparative Analysis of Argonaute-Dependent Small RNA Pathways in Drosophila",

abstract = "The specificity of RNAi pathways is determined by several classes of small RNAs, which include siRNAs, piRNAs, endo-siRNAs, and microRNAs (miRNAs). These small RNAs are invariably incorporated into large Argonaute (Ago)-containing effector complexes known as RNA-induced silencing complexes (RISCs), which they guide to silencing targets. Both genetic and biochemical strategies have yielded conserved molecular components of small RNA biogenesis and effector machineries. However, given the complexity of these pathways, there are likely to be additional components and regulators that remain to be uncovered. We have undertaken a comparative and comprehensive RNAi screen to identify genes that impact three major Ago-dependent small RNA pathways that operate in Drosophila S2 cells. We identify subsets of candidates that act positively or negatively in siRNA, endo-siRNA, and miRNA pathways. Our studies indicate that many components are shared among all three Argonaute-dependent silencing pathways, though each is also impacted by discrete sets of genes.",

keywords = "PROTEINS, RNA",

author = "Rui Zhou and Ikuko Hotta and Denli, {Ahmet M.} and Pengyu Hong and Norbert Perrimon and Hannon, {Gregory J.}",

note = "Funding Information: We thank members of the Hannon and Perrimon labs, especially Sara Cherry for helpful discussion and Julius Brennecke, Ben Czech, and Colin Malone for sharing unpublished data and reagents. We also thank the DRSC staff for technical support, the Bloomington stock center, and Drs. Paul Lasko, Thomas Schwarz, Stephen Stowers, Richard Carthew, and Sara Cherry for fly stocks. We are grateful to Dr. Phillip Sharp for the CXCR4 constructs and to Dr. Paul Lasko for the Bel antibody. We are indebted to Drs. Bernard Mathey-Prevot and Richard Binari for critically reading the manuscript. R.Z. is supported by a Special Fellowship from the Leukemia and Lymphoma Society. This work was supported by grants from the NIH (P.H., N.P., and G.J.H.) and by a kind gift from K.W. Davis (G.J.H.). N.P. and G.J.H. are investigators of the Howard Hughes Medical Institute. ",

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AU - Zhou, Rui

AU - Hotta, Ikuko

AU - Denli, Ahmet M.

AU - Hong, Pengyu

AU - Perrimon, Norbert

AU - Hannon, Gregory J.

N1 - Funding Information: We thank members of the Hannon and Perrimon labs, especially Sara Cherry for helpful discussion and Julius Brennecke, Ben Czech, and Colin Malone for sharing unpublished data and reagents. We also thank the DRSC staff for technical support, the Bloomington stock center, and Drs. Paul Lasko, Thomas Schwarz, Stephen Stowers, Richard Carthew, and Sara Cherry for fly stocks. We are grateful to Dr. Phillip Sharp for the CXCR4 constructs and to Dr. Paul Lasko for the Bel antibody. We are indebted to Drs. Bernard Mathey-Prevot and Richard Binari for critically reading the manuscript. R.Z. is supported by a Special Fellowship from the Leukemia and Lymphoma Society. This work was supported by grants from the NIH (P.H., N.P., and G.J.H.) and by a kind gift from K.W. Davis (G.J.H.). N.P. and G.J.H. are investigators of the Howard Hughes Medical Institute.

PY - 2008/11/21

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N2 - The specificity of RNAi pathways is determined by several classes of small RNAs, which include siRNAs, piRNAs, endo-siRNAs, and microRNAs (miRNAs). These small RNAs are invariably incorporated into large Argonaute (Ago)-containing effector complexes known as RNA-induced silencing complexes (RISCs), which they guide to silencing targets. Both genetic and biochemical strategies have yielded conserved molecular components of small RNA biogenesis and effector machineries. However, given the complexity of these pathways, there are likely to be additional components and regulators that remain to be uncovered. We have undertaken a comparative and comprehensive RNAi screen to identify genes that impact three major Ago-dependent small RNA pathways that operate in Drosophila S2 cells. We identify subsets of candidates that act positively or negatively in siRNA, endo-siRNA, and miRNA pathways. Our studies indicate that many components are shared among all three Argonaute-dependent silencing pathways, though each is also impacted by discrete sets of genes.

AB - The specificity of RNAi pathways is determined by several classes of small RNAs, which include siRNAs, piRNAs, endo-siRNAs, and microRNAs (miRNAs). These small RNAs are invariably incorporated into large Argonaute (Ago)-containing effector complexes known as RNA-induced silencing complexes (RISCs), which they guide to silencing targets. Both genetic and biochemical strategies have yielded conserved molecular components of small RNA biogenesis and effector machineries. However, given the complexity of these pathways, there are likely to be additional components and regulators that remain to be uncovered. We have undertaken a comparative and comprehensive RNAi screen to identify genes that impact three major Ago-dependent small RNA pathways that operate in Drosophila S2 cells. We identify subsets of candidates that act positively or negatively in siRNA, endo-siRNA, and miRNA pathways. Our studies indicate that many components are shared among all three Argonaute-dependent silencing pathways, though each is also impacted by discrete sets of genes.

KW - PROTEINS

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Comparative Analysis of Argonaute-Dependent Small RNA Pathways in Drosophila

Abstract

Keywords

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