부산대학교 도서관

Keywords phase separation; neuronal ribonucleoprotein granule; synaptic new protein synthesis; membraneless organelle; cation-[pi]; arginine methylation; citrullination; frontotemporal dementia; AFM-IR; phase-sensitive fluorescent dyes Highlights * Intermolecular [beta] sheet hydrogen bonding and cation-[pi] interactions drive FUS demixing * Cation-[pi] interactions form between C-terminal arginines and N-terminal tyrosines * Cation-[pi] strength is regulated by arginine methylation and interacting proteins * FUS hypomethylation in FTLD induces FUS gelation and impairs RNP granule function Summary Reversible phase separation underpins the role of FUS in ribonucleoprotein granules and other membrane-free organelles and is, in part, driven by the intrinsically disordered low-complexity (LC) domain of FUS. Here, we report that cooperative cation-[pi] interactions between tyrosines in the LC domain and arginines in structured C-terminal domains also contribute to phase separation. These interactions are modulated by post-translational arginine methylation, wherein arginine hypomethylation strongly promotes phase separation and gelation. Indeed, significant hypomethylation, which occurs in FUS-associated frontotemporal lobar degeneration (FTLD), induces FUS condensation into stable intermolecular [beta]-sheet-rich hydrogels that disrupt RNP granule function and impair new protein synthesis in neuron terminals. We show that transportin acts as a physiological molecular chaperone of FUS in neuron terminals, reducing phase separation and gelation of methylated and hypomethylated FUS and rescuing protein synthesis. These results demonstrate how FUS condensation is physiologically regulated and how perturbations in these mechanisms can lead to disease. Author Affiliation: (1) Cambridge Institute for Medical Research, Department of Clinical Neurosciences, University of Cambridge, Cambridge CB2 0XY, UK (2) Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, UK (3) Department of Physiology, Development, and Neuroscience, University of Cambridge, Cambridge CB2 3DY, UK (4) Tanz Centre for Research in Neurodegenerative Diseases and Departments of Medicine, Medical Biophysics and Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 3H2, Canada (5) Department of Chemical Engineering and Biotechnology, University of Cambridge, Cambridge CB3 0AS, UK (6) Department of Biomedical Sciences, Cedars Sinai Medical Center, Los Angeles, CA 90048, USA (7) Centre for Genomic Regulation, the Barcelona Institute for Science and Technology, 08003 Barcelona, Spain (8) Universitat Pompeu Fabra, 08003 Barcelona, Spain (9) Institucio Catalana de Recerca i Estudis Avancats, 08010 Barcelona, Spain (10) Departments of Medicine, Neurology, and Ophthalmology and Departments of Epidemiology and Biostatistics, Boston University, Boston, MA 02118, USA (11) Cavendish Laboratory, Department of Physics, University of Cambridge, Cambridge CB3 0HE, UK * Corresponding author Article History: Received 25 June 2017; Revised 11 December 2017; Accepted 21 March 2018 (miscellaneous) Published: April 19, 2018 (footnote)12 These authors contributed equally (footnote)13 Lead Contact Byline: Seema Qamar (1,12), GuoZhen Wang (1,12), Suzanne J. Randle (1,12), Francesco Simone Ruggeri (2,12), Juan A. Varela (2,12), Julie Qiaojin Lin (3,12), Emma C. Phillips (1,12), Akinori Miyashita (4), Declan Williams (4), Florian Strohl (5), William Meadows (1), Rodylyn Ferry (4), Victoria J. Dardov (6), Gian G. Tartaglia (7,8,9), Lindsay A. Farrer (10), Gabriele S. Kaminski Schierle (5), Clemens F. Kaminski (5), Christine E. Holt (3), Paul E. Fraser (4), Gerold Schmitt-Ulms (4), David Klenerman (2), Tuomas Knowles (2,11), Michele Vendruscolo (2), Peter St George-Hyslop [p.hyslop@utoronto.ca] (1,4,13,*)