title: FUS functions in coupling transcription to splicing by mediating an interaction between RNAP II and U1 snRNP.
abstract: Pre-mRNA splicing is coupled to transcription by RNA polymerase II (RNAP II). We previously showed that U1 small nuclear ribonucleoprotein (snRNP) associates with RNAP II, and both RNAP II and U1 snRNP are also the most abundant factors associated with the protein fused-in-sarcoma (FUS), which is mutated to cause the neurodegenerative disease amyotrophic lateral sclerosis. Here, we show that an antisense morpholino that base-pairs to the 5' end of U1 snRNA blocks splicing in the coupled system and completely disrupts the association between U1 snRNP and both FUS and RNAP II, but has no effect on the association between FUS and RNAP II. Conversely, we found that U1 snRNP does not interact with RNAP II in FUS knockdown extracts. Moreover, using these extracts, we found that FUS must be present during the transcription reaction in order for splicing to occur. Together, our data lead to a model that FUS functions in coupling transcription to splicing via mediating an interaction between RNAP II and U1 snRNP.
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, MA 02115.
authors.forename: Yong
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, MA 02115 robin_reed@hms.harvard.edu.
authors.forename: Robin
authors.initials: R
authors.lastname: Reed
mesh_terms: D001483:Base Sequence; D006801:Humans; D012319:RNA Polymerase II / Q000378:metabolism*; D012326:RNA Splicing / Q000502:physiology*; D012342:RNA, Small Nuclear / Q000737:chemistry / Q000378:metabolism; D034702:RNA-Binding Protein FUS / Q000378:metabolism / Q000502:physiology*; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000378:metabolism*; D014158:Transcription, Genetic / Q000502:physiology*
chemical_list: C000599550:FUS protein, human; D012342:RNA, Small Nuclear; D034702:RNA-Binding Protein FUS; D017412:Ribonucleoprotein, U1 Small Nuclear; C405483:U1 small nuclear RNA; D012319:RNA Polymerase II
keywords: ALS; RNA polymerase II; U1 snRNP; coupling transcription to splicing