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Ataxin-2, Twenty-four and Dicer-2 are components of a non-canonical cytoplasmic polyadenylation complex

ABSTRACT: Cytoplasmic polyadenylation is a mechanism to promote mRNA translation in a wide variety of biological contexts. A canonical complex centered around the conserved RNA-binding protein family CPEB has been shown to be responsible for this process. We have previously reported evidence for an alternative non-canonical, CPEB-independent complex in Drosophila, of which the RNA-interference factor Dicer-2 is a component. Here, we investigate Dicer-2 mRNA targets and protein co-factors in cytoplasmic polyadenylation. Using RIP-Seq analysis we identify hundreds of novel Dicer-2 target transcripts, ~50% of which were previously found as targets of the cytoplasmic poly(A) polymerase Wispy, suggesting widespread roles of Dicer-2 in cytoplasmic polyadenylation. Large-scale immunoprecipitation revealed Ataxin-2 and Twenty-four among the high-confidence interactors of Dicer-2. Functional analysis indicate that both factors form an RNA-independent complex with Dicer-2, and are required for cytoplasmic polyadenylation of Dicer-2 targets. Our results reveal the composition of a novel cytoplasmic polyadenylation complex that operates during Drosophila early embryogenesis.

ORGANISM(S): Drosophila melanogaster

PROVIDER: GSE189868 | GEO | 2022/08/29

REPOSITORIES: GEO

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Drosophila early embryos LC-MSMS

Project description:Cytoplasmic polyadenylation is a mechanism to promote mRNA translation in a wide variety of biological contexts.The conserved RNA-binding protein family CPEB has been shown to mediate canonical cytoplasmic polyadenylation of target transcripts. We have previously reported evidence for RNA-interference factor Dicer-2 as a component of a non-canonical complex, that operates independent of CPEB in Drosophila. In this study, we investigate Dicer-2 mRNA targets and protein co-factors in cytoplasmic polyadenylation. Using RIP‐Seq analysis we identify hundreds of potential Dicer-2 target transcripts, ~60% of which were previously found as targets of the cytoplasmic poly(A) polymerase Wispy, suggesting widespread roles of Dicer-2 in cytoplasmic polyadenylation. Large-scale immunoprecipitation and mass spectrometry revealed Ataxin-2 and Twenty-four among the high-confidence interactors of Dicer-2. Complex analyses indicated that both factors form an RNA‐independent complex with Dicer‐2, and mediate interactions of Dicer‐2 with Wispy. Functional poly(A)‐test analyses showed that Twenty‐four and Ataxin-2 are required for cytoplasmic polyadenylation of a subset of Dicer‐2 targets. Our results reveal components of a novel cytoplasmic polyadenylation complex that operates during Drosophila early embryogenesis.

2022-08-30 | PXD030281 | Pride

Comparative analyses of vertebrate CPEB-family of proteins define two sub-families with distinct, but coordinated, functions in post-transcriptional regulation of gene expressionComparative analyses of vertebrate CPEB-family of proteins define two sub-families with distinct, but coordinated, functions in post-transcriptional regulation of gene expression

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2022-10-05 | PXD030480 | Pride

Comparative analyses of vertebrate CPEB-family of proteins define two sub-families with distinct, but coordinated, functions in post-transcriptional regulation of gene expression

2022-10-05 | PXD033349 | Pride

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2017-07-17 | GSE68098 | GEO

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2016-10-30 | GSE59611 | GEO

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2016-05-18 | GSE81495 | GEO