Cell-free formation of RNA granules: Bound RNAs identify features and components of cellular assemblies

Tina W. Han, Masato Kato, Shanhai Xie, Leeju C. Wu, Hamid Mirzaei, Jimin Pei, Min Chen, Yang Xie, Jeffrey Allen, Guanghua Xiao, Steven L. McKnight

Research output: Contribution to journalArticle

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Abstract

Cellular granules lacking boundary membranes harbor RNAs and their associated proteins and play diverse roles controlling the timing and location of protein synthesis. Formation of such granules was emulated by treatment of mouse brain extracts and human cell lysates with a biotinylated isoxazole (b-isox) chemical. Deep sequencing of the associated RNAs revealed an enrichment for mRNAs known to be recruited to neuronal granules used for dendritic transport and localized translation at synapses. Precipitated mRNAs contain extended 3′ UTR sequences and an enrichment in binding sites for known granule-associated proteins. Hydrogels composed of the low complexity (LC) sequence domain of FUS recruited and retained the same mRNAs as were selectively precipitated by the b-isox chemical. Phosphorylation of the LC domain of FUS prevented hydrogel retention, offering a conceptual means of dynamic, signal-dependent control of RNA granule assembly. PaperClip:

Original languageEnglish (US)
Pages (from-to)768-779
Number of pages12
JournalCell
Volume149
Issue number4
DOIs
Publication statusPublished - May 11 2012

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ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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