Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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RNAi knock down of mRNA decapping co-activators in Drosophila S2 cells

ABSTRACT: We analyzed mRNA expression profiles in Drosophila melanogaster S2 cells that had been depleted of proteins known as mRNA decapping co-activators. mRNA decapping is catalyzed by DCP2, and DCP2 activity is stimulated by decapping co-activators. This group of proteins includes DCP1, Hedls (also known as Ge-1), LSm16 (also known as EDC3), rck/p54 (also known as DHH1 or Me31B), Pat1, and the heptameric LSm1-7 complex. We used the RNA interference technology to deplete cultured S2 cells of DCP1 (CG11183), Ge-1 (CG6181), Pat1 (CG5208), LSm16 (CG6311), and LSm1 (CG4279). We used Affymetrix oligonucleotide microarrays to analyze two independent samples for each depletion. We included the following controls: “mock” RNAi treatment and GFP dsRNA treatment (two profiles each). We also profiled AGO1 (CG6671) depleted cells (3 independent samples). AGO1 is a key protein required for miRNA-mediated gene silencing. We had shown previously that silencing by miRNAs involves decapping of target mRNAs.

INSTRUMENT(S): 418 [Affymetrix]

ORGANISM(S): Drosophila melanogaster

SUBMITTER: Jan Rehwinkel 

PROVIDER: E-MEXP-1012 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Target-specific requirements for enhancers of decapping in miRNA-mediated gene silencing.

Eulalio Ana A   Rehwinkel Jan J   Stricker Mona M   Huntzinger Eric E   Yang Schu-Fee SF   Doerks Tobias T   Dorner Silke S   Bork Peer P   Boutros Michael M   Izaurralde Elisa E  

Genes & development 20070927 20

microRNAs (miRNAs) silence gene expression by suppressing protein production and/or by promoting mRNA decay. To elucidate how silencing is accomplished, we screened an RNA interference library for suppressors of miRNA-mediated regulation in Drosophila melanogaster cells. In addition to proteins known to be required for miRNA biogenesis and function (i.e., Drosha, Pasha, Dicer-1, AGO1, and GW182), the screen identified the decapping activator Ge-1 as being required for silencing by miRNAs. Deplet  ...[more]

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