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P-bodies are sites of rapid RNA decay during the neural crest epithelial-mesenchymal transition

Hutchins, Erica J. and Piacentino, Michael L. and Bronner, Marianne E. (2020) P-bodies are sites of rapid RNA decay during the neural crest epithelial-mesenchymal transition. . (Unpublished)

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The epithelial-mesenchymal transition (EMT) drives cellular movements during development to create specialized tissues and structures in metazoans, using mechanisms often coopted during metastasis. Neural crest cells are a multipotent stem cell population that undergo a developmentally regulated EMT and are prone to metastasis in the adult, providing an excellent model to study cell state changes and mechanisms underlying EMT. A hallmark of neural crest EMT during avian development is temporally restricted expression followed by rapid down-regulation of the Wnt antagonist Draxin. Using live RNA imaging, here we demonstrate that rapid clearance of Draxin transcripts is mediated post-transcriptionally via localization to processing bodies (P-bodies), small cytoplasmic granules which are established sites of RNA processing. Contrasting with recent work in immortalized cell lines suggesting that P-bodies are sites of storage rather than degradation, we show that targeted decay of Draxin occurs within P-bodies during neural crest migration. Furthermore, P-body disruption via DDX6 knockdown inhibits not only endogenous Draxin down-regulation but also neural crest EMT in vivo. Together, our data highlight a novel and important role for P-bodies in an intact organismal context−controlling a developmental EMT program via post-transcriptional target degradation.

Item Type:Report or Paper (Discussion Paper)
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URLURL TypeDescription Paper ItemCode
Hutchins, Erica J.0000-0002-4316-0333
Piacentino, Michael L.0000-0003-1773-031X
Bronner, Marianne E.0000-0003-4274-1862
Additional Information:The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license. Posted August 01, 2020. We thank A. Collazo and G. Spigolon for imaging assistance at the Caltech Biological Imaging Facility; M. Schwarzkopf and G. Shin (Molecular Technologies) for HCR probe design; S. Manohar, G. da Silva Pescador, and C.J. Andrews for cloning assistance; R. Galton for pilot HCR experiments; and R. Singer, J. Chao, and E. Izaurralde for essential reagents made available via Addgene. This work was supported by the National Institutes of Health [R01DE027538 and R01DE027568 to M.E.B; K99DE028592 to E.J.H; K99DE029240 to M.L.P.]. Data availability: All data generated and/or analyzed during the current study are available from the corresponding author on reasonable request. Code availability: Information for accessing the JaCoP plugin used to calculate Mander’s coefficients is available: Author Contributions: Project was conceived by E.J.H. and M.E.B. Experimental design and data interpretation were conducted by E.J.H, M.L.P, and M.E.B. Time-lapse experiments were performed by E.J.H. and M.L.P. Chick electroporations and explants were performed by E.J.H. Constructs were designed and generated by E.J.H. and M.L.P. Embryology, hybridization chain reaction experiments, imaging, and quantitation and statistical analyses were performed by E.J.H. Manuscript was written by E.J.H. and M.E.B., with editing by M.L.P. The authors have declared no competing interest.
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Record Number:CaltechAUTHORS:20200803-123033784
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Official Citation:P-bodies are sites of rapid RNA decay during the neural crest epithelial—mesenchymal transition. Erica J. Hutchins, Michael L. Piacentino, Marianne E. Bronner. bioRxiv 2020.07.31.231860; doi:
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:104703
Deposited By: Tony Diaz
Deposited On:03 Aug 2020 20:23
Last Modified:16 Nov 2021 18:34

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