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PMID: 18023351 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

A whole-genome RNAi Screen for C. elegans miRNA pathway genes.

Current biology : CB ·Vol. 17 ·No. 23 ·2007-12-04 ·Pages 2013-22

Parry DH, Xu J, Ruvkun G

Abstract

miRNAs are an abundant class of small, endogenous regulatory RNAs. Although it is now appreciated that miRNAs are involved in a broad range of biological processes, relatively little is known about the actual mechanism by which miRNAs downregulate target gene expression. An exploration of which protein cofactors are necessary for a miRNA to downregulate a target gene should reveal more fully the molecular mechanisms by which miRNAs are processed, trafficked, and regulate their target genes. A weak allele of the C. elegans miRNA gene let-7 was used as a sensitized genetic background for a whole-genome RNAi screen to detect miRNA pathway genes, and 213 candidate miRNA pathway genes were identified. About 2/3 of the 61 candidates with the strongest phenotype were validated through genetic tests examining the dependence of the let-7 phenotype on target genes known to function in the let-7 pathway. Biochemical tests for let-7 miRNA production place the function of nearly all of these new miRNA pathway genes downstream of let-7 expression and processing. By monitoring the downregulation of the protein product of the lin-14 mRNA, which is the target of the lin-4 miRNA, we have identified 19 general miRNA pathway genes. The 213 candidate miRNA pathway genes identified could act at steps that produce and traffic miRNAs or in downstream steps that detect miRNA::mRNA duplexes to regulate mRNA translation. The 19 validated general miRNA pathway genes are good candidates for genes that may define protein cofactors for sorting or targeting miRNA::mRNA duplexes, or for recognizing the miRNA base-paired to the target mRNA to downregulate translation.

MeSH Terms
Animals Caenorhabditis elegans/genetics,growth & development,metabolism Caenorhabditis elegans Proteins/genetics,metabolism Computational Biology Gene Expression Regulation Genome, Helminth MicroRNAs/genetics,metabolism Nuclear Proteins/genetics,metabolism RNA Interference RNA, Helminth/genetics,metabolism RNA, Messenger/genetics,metabolism RNA, Small Interfering/genetics,metabolism
Chemicals
Caenorhabditis elegans Proteins LIN-14 protein, C elegans MicroRNAs Nuclear Proteins RNA, Helminth RNA, Messenger RNA, Small Interfering let-7 microRNA, C elegans
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Parry Devin H
Department of Genetics, Harvard Medical School, Richard B. Simches Research Building, 185 Cambridge Street, CPZN-7250, Boston, Massachusetts 02114-2790, USA.
Xu Jinling
Ruvkun Gary
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Article Info
Journal
Current biology : CB
Abbr.
Curr Biol
ISSN
0960-9822
DOI
10.1016/j.cub.2007.10.058
Published
2007-12-04
Epub
2007-00-20
Pages
2013-22
Language
English
Region
England
NLM ID
9107782
PMCID
PMC2211719
Subset
IM
Grants
NIGMS NIH HHS · R01 GM044619 · United States
NIGMS NIH HHS · R01 GM044619-16 · United States
NIGMS NIH HHS · R01-GM44619 · United States
External Links
PubMed source DOI: 10.1016/j.cub.2007.10.058 PMC Full Text View journal details
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