title: An RRM-ZnF RNA recognition module targets RBM10 to exonic sequences to promote exon exclusion.
abstract: RBM10 is an RNA-binding protein that plays an essential role in development and is frequently mutated in the context of human disease. RBM10 recognizes a diverse set of RNA motifs in introns and exons and regulates alternative splicing. However, the molecular mechanisms underlying this seemingly relaxed sequence specificity are not understood and functional studies have focused on 3΄ intronic sites only. Here, we dissect the RNA code recognized by RBM10 and relate it to the splicing regulatory function of this protein. We show that a two-domain RRM1-ZnF unit recognizes a GGA-centered motif enriched in RBM10 exonic sites with high affinity and specificity and test that the interaction with these exonic sequences promotes exon skipping. Importantly, a second RRM domain (RRM2) of RBM10 recognizes a C-rich sequence, which explains its known interaction with the intronic 3΄ site of NUMB exon 9 contributing to regulation of the Notch pathway in cancer. Together, these findings explain RBM10's broad RNA specificity and suggest that RBM10 functions as a splicing regulator using two RNA-binding units with different specificities to promote exon skipping.
authors.affiliation: Institute of Structural and Molecular Biology, University College London, London, WC1E 6BT, UK.
authors.forename: Katherine M
authors.initials: KM
authors.lastname: Collins
authors.affiliation: Centre for Developmental Neurobiology, King's College London, London SE1 1UL, UK.
authors.forename: Yaroslav A
authors.initials: YA
authors.lastname: Kainov
authors.affiliation: Structural Biology Science Technology Platform, The Francis Crick Institute, London NW1 1AT, UK.
authors.forename: Evangelos
authors.initials: E
authors.lastname: Christodolou
authors.affiliation: Donnelly Centre, University of Toronto, Toronto M5S 3E1, Canada.
authors.forename: Debashish
authors.initials: D
authors.lastname: Ray
authors.affiliation: Donnelly Centre, University of Toronto, Toronto M5S 3E1, Canada.
authors.forename: Quaid
authors.initials: Q
authors.lastname: Morris
authors.affiliation: Donnelly Centre, University of Toronto, Toronto M5S 3E1, Canada.
authors.forename: Timothy
authors.initials: T
authors.lastname: Hughes
authors.affiliation: Macromolecular Structure Laboratory, The Francis Crick Institute, London NW1 1AT, UK.
authors.forename: Ian A
authors.initials: IA
authors.lastname: Taylor
authors.affiliation: Centre for Developmental Neurobiology, King's College London, London SE1 1UL, UK.
authors.forename: Eugene V
authors.initials: EV
authors.lastname: Makeyev
authors.affiliation: Institute of Structural and Molecular Biology, University College London, London, WC1E 6BT, UK.
authors.forename: Andres
authors.initials: A
authors.lastname: Ramos
mesh_terms: D001324:Autoantigens; D001483:Base Sequence; D001665:Binding Sites; D005091:Exons; D057809:HEK293 Cells; D006801:Humans; D011485:Protein Binding; D012326:RNA Splicing; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000502:physiology*; D016335:Zinc Fingers
chemical_list: D001324:Autoantigens; C512260:RBM10 protein, human; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C465663:TNRC6A protein, human