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Status |
Public on Aug 29, 2022 |
Title |
Ataxin-2, Twenty-four and Dicer-2 are components of a non-canonical cytoplasmic polyadenylation complex |
Organism |
Drosophila melanogaster |
Experiment type |
Expression profiling by high throughput sequencing
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Summary |
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.
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Overall design |
RNA immunoprecipitation-sequencing (RIP-Seq) was performed to indentify Dicer-2 RNA targets in Drosophila melanogaster early embryo extracts. Three types of samples, in sextuplicates, were produced: (1) RNA eluate from specific immunoprecipitation against Dicer-2, either compared to (2) RNA from non-specific IgG immunoprecipitation (control) or (3) RNA from Input samples.
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Contributor(s) |
Nadimpalli HP, Guitart T, Gebauer F |
Citation(s) |
36114004 |
Submission date |
Nov 30, 2021 |
Last update date |
Nov 28, 2022 |
Contact name |
Estefania Mancini |
E-mail(s) |
estefania.mancini@crg.eu
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Phone |
34693564066
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Organization name |
Centre for Genomic Regulation
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Department |
GENE REGULATION, STEM CELLS AND CANCER
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Lab |
Regulation of Alternative pre-mRNA Splicing during Cell Differentiation, Development and Disease
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Street address |
C/ Dr. Aiguader, 88
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City |
Barcelona |
State/province |
Barcelona |
ZIP/Postal code |
08003 |
Country |
Spain |
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Platforms (1) |
GPL17275 |
Illumina HiSeq 2500 (Drosophila melanogaster) |
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Samples (17)
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Relations |
BioProject |
PRJNA784931 |
SRA |
SRP348641 |