title: A loss of FUS/TLS function leads to impaired cellular proliferation.
abstract: Fused in sarcoma/translocated in liposarcoma (FUS/TLS or FUS) is a multifunctional RNA/DNA-binding protein that is pathologically associated with cancer and neurodegeneration. To gain insight into the vital functions of FUS and how a loss of FUS function impacts cellular homeostasis, FUS expression was reduced in different cellular models through RNA interference. Our results show that a loss of FUS expression severely impairs cellular proliferation and leads to an increase in phosphorylated histone H3, a marker of mitotic arrest. A quantitative proteomics analysis performed on cells undergoing various degrees of FUS knockdown revealed protein expression changes for known RNA targets of FUS, consistent with a loss of FUS function with respect to RNA processing. Proteins that changed in expression as a function of FUS knockdown were associated with multiple processes, some of which influence cell proliferation including cell cycle regulation, cytoskeletal organization, oxidative stress and energy homeostasis. FUS knockdown also correlated with increased expression of the closely related protein EWS (Ewing's sarcoma). We demonstrate that the maladaptive phenotype resulting from FUS knockdown is reversible and can be rescued by re-expression of FUS or partially rescued by the small-molecule rolipram. These results provide insight into the pathways and processes that are regulated by FUS, as well as the cellular consequences for a loss of FUS function.
authors.affiliation: Department of Neurology, University of Massachusetts Medical School, Albert Sherman Center, Worcester, MA, USA.
authors.forename: C L
authors.initials: CL
authors.lastname: Ward
authors.affiliation: Department of Neurology, University of Massachusetts Medical School, Albert Sherman Center, Worcester, MA, USA.
authors.forename: K J
authors.initials: KJ
authors.lastname: Boggio
authors.affiliation: Laboratory for Drug Discovery in Neurodegeneration, Harvard NeuroDiscovery Center, Brigham and Women's Hospital, Harvard Medical School, Cambridge, MA, USA.
authors.forename: B N
authors.initials: BN
authors.lastname: Johnson
authors.affiliation: Laboratory for Drug Discovery in Neurodegeneration, Harvard NeuroDiscovery Center, Brigham and Women's Hospital, Harvard Medical School, Cambridge, MA, USA.
authors.forename: J B
authors.initials: JB
authors.lastname: Boyd
authors.affiliation: Department of Cell and Developmental Biology, University of Massachusetts Medical School, Worcester, MA, USA.
authors.forename: S
authors.initials: S
authors.lastname: Douthwright
authors.affiliation: 1] Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA, USA [2] Proteomics and Mass Spectrometry Facility, University of Massachusetts Medical School, Shrewsbury, MA, USA.
authors.forename: S A
authors.initials: SA
authors.lastname: Shaffer
authors.affiliation: Department of Neurology, University of Massachusetts Medical School, Albert Sherman Center, Worcester, MA, USA.
authors.forename: J E
authors.initials: JE
authors.lastname: Landers
authors.affiliation: Laboratory for Drug Discovery in Neurodegeneration, Harvard NeuroDiscovery Center, Brigham and Women's Hospital, Harvard Medical School, Cambridge, MA, USA.
authors.forename: M A
authors.initials: MA
authors.lastname: Glicksman
authors.affiliation: 1] Department of Neurology, University of Massachusetts Medical School, Albert Sherman Center, Worcester, MA, USA [2] Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA, USA.
authors.forename: D A
authors.initials: DA
authors.lastname: Bosco
mesh_terms: D002460:Cell Line; D049109:Cell Proliferation; D002477:Cells / Q000166:cytology* / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006657:Histones / Q000378:metabolism; D006801:Humans; D059566:M Phase Cell Cycle Checkpoints; D010766:Phosphorylation; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D034702:RNA-Binding Protein FUS / Q000172:deficiency* / Q000235:genetics
chemical_list: D006657:Histones; D034741:RNA, Small Interfering; D034702:RNA-Binding Protein FUS