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PMID: 26054396 Published · epublish English Journal Article Research Support, Non-U.S. Gov't

Drosophila Imp iCLIP identifies an RNA assemblage coordinating F-actin formation.

Genome biology ·Vol. 16 ·2015-06-09 ·Pages 123

Hansen HT, Rasmussen SH, Adolph SK, Plass M, Krogh A, Sanford J, Nielsen FC, Christiansen J

Abstract

Post-transcriptional RNA regulons ensure coordinated expression of monocistronic mRNAs encoding functionally related proteins. In this study, we employ a combination of RIP-seq and short- and long-wave individual-nucleotide resolution crosslinking and immunoprecipitation (iCLIP) technologies in Drosophila cells to identify transcripts associated with cytoplasmic ribonucleoproteins (RNPs) containing the RNA-binding protein Imp. We find extensive binding of Imp to 3' UTRs of transcripts that are involved in F-actin formation. A common denominator of the RNA-protein interface is the presence of multiple motifs with a central UA-rich element flanked by CA-rich elements. Experiments in single cells and intact flies reveal compromised actin cytoskeletal dynamics associated with low Imp levels. The former shows reduced F-actin formation and the latter exhibits abnormal neuronal patterning. This demonstrates a physiological significance of the defined RNA regulon. Our data imply that Drosophila Imp RNPs may function as cytoplasmic mRNA assemblages that encode proteins which participate in actin cytoskeletal remodeling. Thus, they may facilitate coordinated protein expression in sub-cytoplasmic locations such as growth cones.

MeSH Terms
3' Untranslated Regions Actin Cytoskeleton/metabolism Actins/metabolism Animals Cells, Cultured Cytoplasmic Granules/chemistry Drosophila/embryology,genetics,metabolism Drosophila Proteins/analysis,antagonists & inhibitors,metabolism High-Throughput Nucleotide Sequencing Immunoprecipitation Male Nervous System/embryology RNA, Messenger/analysis,metabolism RNA-Binding Proteins/analysis,antagonists & inhibitors,metabolism Ribonucleoproteins/metabolism Sequence Analysis, RNA
Chemicals
3' Untranslated Regions Actins Drosophila Proteins Imp protein, Drosophila RNA, Messenger RNA-Binding Proteins Ribonucleoproteins
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Hansen Heidi Theil
Department of Biology, Center for Computational and Applied Transcriptomics, University of Copenhagen, Ole Maaloes Vej 5, 2200, Copenhagen, Denmark. hhansen@bio.ku.dk.
Rasmussen Simon Horskjær
Department of Biology, Center for Computational and Applied Transcriptomics, University of Copenhagen, Ole Maaloes Vej 5, 2200, Copenhagen, Denmark. simras@binf.ku.dk.
Adolph Sidsel Kramshøj
Department of Biology, Center for Computational and Applied Transcriptomics, University of Copenhagen, Ole Maaloes Vej 5, 2200, Copenhagen, Denmark. sidselka@yahoo.dk.
Plass Mireya
Department of Biology, Center for Computational and Applied Transcriptomics, University of Copenhagen, Ole Maaloes Vej 5, 2200, Copenhagen, Denmark. mplass@binf.ku.dk.
Krogh Anders
Department of Biology, Center for Computational and Applied Transcriptomics, University of Copenhagen, Ole Maaloes Vej 5, 2200, Copenhagen, Denmark. akrogh@bio.ku.dk.
Sanford Jeremy
MCD Biology, University of California, Santa Cruz, CA, 95064, USA. jsanfor2@ucsc.edu.
Nielsen Finn Cilius
Center for Genomic Medicine, Rigshospitalet, University of Copenhagen, 2100, Copenhagen, Denmark. cilius@me.com.
Christiansen Jan
Department of Biology, Center for Computational and Applied Transcriptomics, University of Copenhagen, Ole Maaloes Vej 5, 2200, Copenhagen, Denmark. janchr@bio.ku.dk.
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Article Info
Journal
Genome biology
Abbr.
Genome Biol
ISSN
1474-760X
Published
2015-06-09
Epub
2015-00-09
Pages
123
Language
English
Region
England
NLM ID
100960660
PMCID
PMC4477473
Subset
IM
Grants
NIGMS NIH HHS · R01 GM109146 · United States
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