title: Loss of MBNL leads to disruption of developmentally regulated alternative polyadenylation in RNA-mediated disease.
abstract: Inhibition of muscleblind-like (MBNL) activity due to sequestration by microsatellite expansion RNAs is a major pathogenic event in the RNA-mediated disease myotonic dystrophy (DM). Although MBNL1 and MBNL2 bind to nascent transcripts to regulate alternative splicing during muscle and brain development, another major binding site for the MBNL protein family is the 3' untranslated region of target RNAs. Here, we report that depletion of Mbnl proteins in mouse embryo fibroblasts leads to misregulation of thousands of alternative polyadenylation events. HITS-CLIP and minigene reporter analyses indicate that these polyadenylation switches are a direct consequence of MBNL binding to target RNAs. Misregulated alternative polyadenylation also occurs in skeletal muscle in a mouse polyCUG model and human DM, resulting in the persistence of neonatal polyadenylation patterns. These findings reveal an additional developmental function for MBNL proteins and demonstrate that DM is characterized by misregulation of pre-mRNA processing at multiple levels.
authors.affiliation: Department of Molecular Genetics and Microbiology, Center for NeuroGenetics and the Genetics Institute, University of Florida, College of Medicine, Gainesville, FL 32610, USA.
authors.forename: Ranjan
authors.initials: R
authors.lastname: Batra
authors.affiliation: Department of Molecular Genetics and Microbiology, Center for NeuroGenetics and the Genetics Institute, University of Florida, College of Medicine, Gainesville, FL 32610, USA.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Charizanis
authors.affiliation: Department of Molecular Genetics and Microbiology, Center for NeuroGenetics and the Genetics Institute, University of Florida, College of Medicine, Gainesville, FL 32610, USA.
authors.forename: Mini
authors.initials: M
authors.lastname: Manchanda
authors.affiliation: Department of Molecular Genetics and Microbiology, Center for NeuroGenetics and the Genetics Institute, University of Florida, College of Medicine, Gainesville, FL 32610, USA.
authors.forename: Apoorva
authors.initials: A
authors.lastname: Mohan
authors.affiliation: Department of Molecular Genetics and Microbiology, Center for NeuroGenetics and the Genetics Institute, University of Florida, College of Medicine, Gainesville, FL 32610, USA.
authors.forename: Moyi
authors.initials: M
authors.lastname: Li
authors.affiliation: Department of Molecular Genetics and Microbiology, Center for NeuroGenetics and the Genetics Institute, University of Florida, College of Medicine, Gainesville, FL 32610, USA.
authors.forename: Dustin J
authors.initials: DJ
authors.lastname: Finn
authors.affiliation: Department of Molecular Genetics and Microbiology, Center for NeuroGenetics and the Genetics Institute, University of Florida, College of Medicine, Gainesville, FL 32610, USA.
authors.forename: Marianne
authors.initials: M
authors.lastname: Goodwin
authors.affiliation: Department of Systems Biology, Department of Biochemistry and Molecular Biophysics, Center for Motor Neuron Biology and Disease, Columbia University, New York, NY 10032, USA.
authors.forename: Chaolin
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Department of Neurology, University of Rochester Medical Center, Rochester, NY 14642, USA; Department of Gene Expression, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University, Umultowska 89, 61-614 Poznan, Poland.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Sobczak
authors.affiliation: Department of Neurology, University of Rochester Medical Center, Rochester, NY 14642, USA.
authors.forename: Charles A
authors.initials: CA
authors.lastname: Thornton
authors.affiliation: Department of Molecular Genetics and Microbiology, Center for NeuroGenetics and the Genetics Institute, University of Florida, College of Medicine, Gainesville, FL 32610, USA. Electronic address: mswanson@ufl.edu.
authors.forename: Maurice S
authors.initials: MS
authors.lastname: Swanson
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D017398:Alternative Splicing / Q000235:genetics*; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D051379:Mice; D018895:Microsatellite Repeats / Q000235:genetics; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism; D009223:Myotonic Dystrophy / Q000235:genetics; D026723:Polyadenylation / Q000235:genetics*; D011485:Protein Binding; D034622:RNA Interference; D012322:RNA Precursors / Q000235:genetics; D012323:RNA Processing, Post-Transcriptional / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D020413:3' Untranslated Regions; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; C522920:MBNL3 protein, mouse; C479922:Mbnl1 protein, mouse; C529512:Mbnl2 protein, mouse; D012322:RNA Precursors; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins