title: Cell-free formation of RNA granules: bound RNAs identify features and components of cellular assemblies.
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.
authors.affiliation: Department of Biochemistry, UT Southwestern Medical Center, Dallas, TX 75390-9152, USA.
authors.forename: Tina W
authors.initials: TW
authors.lastname: Han
authors.forename: Masato
authors.initials: M
authors.lastname: Kato
authors.forename: Shanhai
authors.initials: S
authors.lastname: Xie
authors.forename: Leeju C
authors.initials: LC
authors.lastname: Wu
authors.forename: Hamid
authors.initials: H
authors.lastname: Mirzaei
authors.forename: Jimin
authors.initials: J
authors.lastname: Pei
authors.forename: Min
authors.initials: M
authors.lastname: Chen
authors.forename: Yang
authors.initials: Y
authors.lastname: Xie
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Allen
authors.forename: Guanghua
authors.initials: G
authors.lastname: Xiao
authors.forename: Steven L
authors.initials: SL
authors.lastname: McKnight
mesh_terms: D000818:Animals; D019921:Biotinylation; D001921:Brain / Q000166:cytology* / Q000378:metabolism; D002460:Cell Line; D002474:Cell-Free System; D006801:Humans; D007555:Isoxazoles / Q000378:metabolism; D051379:Mice; D012313:RNA / Q000032:analysis* / Q000378:metabolism*; D034443:RNA Transport; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D012261:Ribonucleoproteins / Q000737:chemistry*
chemical_list: D007555:Isoxazoles; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; D012313:RNA