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bioRxiv · 10.1101/2022.11.23.517728

The RNA Binding proteome of axonal mRNAs in sympathetic neurons

Luisier, R.·Andreassi, C.·Riccio, A.
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Abstract

BackgroundNeurons are morphologically complex cells that rely on the compartmentalization of protein expression to develop and maintain their cytoarchitecture. Targeting of RNA transcripts to axons is one of the mechanisms that allows rapid local translation of proteins in response to extracellular signals. 3 untranslated regions (UTRs) of mRNA are non-coding sequences that play a critical role in determining transcript localisation and translation by interacting with specific RNA binding proteins (RBPs). However, how 3UTRs contribute to mRNA metabolism and the nature of RBP complexes responsible for these functions remain elusive. ResultsWe performed 3 end sequencing of RNA isolated from axons and cell bodies of sympathetic neurons exposed to either Nerve Growth factor (NGF) or Neurotrophin 3 (NT3). NGF and NT3 are growth factors essential for sympathetic neuron development that act through distinct signalling mechanisms. Whereas NT3 is thought to act only locally, NGF signals back from axons to the cell bodies. We discovered that both NGF and NT3 affect transcription and alternative polyadenylation and induce the localisation of specific 3UTR isoforms to axons. The finding that many transcripts with short 3UTR were detected only in axons suggest that these may undergo local post-transcriptional remodelling. The integration of our data with CLIP-sequencing data revealed that long 3UTR isoforms associate with RBP complexes in the nucleus, and once in axons, regulate cytoplasmic 3 UTR isoform cleavage into shorter isoform. ConclusionsOur findings shed new light on the complex interplay between nuclear polyadenylation, mRNA localisation and local 3UTR remodelling in developing neurons. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=106 SRC="FIGDIR/small/517728v1_ufig1.gif" ALT="Figure 1"> View larger version (36K): org.highwire.dtl.DTLVardef@17196eforg.highwire.dtl.DTLVardef@6c26adorg.highwire.dtl.DTLVardef@5f335borg.highwire.dtl.DTLVardef@24a6d6_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Luisier, R., Andreassi, C., Riccio, A.. 2022-11-24. The RNA Binding proteome of axonal mRNAs in sympathetic neurons. https://doi.org/10.1101/2022.11.23.517728

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