title: The splicing factor FUBP1 is required for the efficient splicing of oncogene MDM2 pre-mRNA.
abstract: Alternative splicing of the oncogene MDM2 is a phenomenon that occurs in cells in response to genotoxic stress and is also a hallmark of several cancer types with important implications in carcinogenesis. However, the mechanisms regulating this splicing event remain unclear. Previously, we uncovered the importance of intron 11 in MDM2 that affects the splicing of a damage-responsive MDM2 minigene. Here, we have identified discrete cis regulatory elements within intron 11 and report the binding of FUBP1 (Far Upstream element-Binding Protein 1) to these elements and the role it plays in MDM2 splicing. Best known for its oncogenic role as a transcription factor in the context of c-MYC, FUBP1 was recently described as a splicing regulator with splicing repressive functions. In the case of MDM2, we describe FUBP1 as a positive splicing regulatory factor. We observed that blocking the function of FUBP1 in in vitro splicing reactions caused a decrease in splicing efficiency of the introns of the MDM2 minigene. Moreover, knockdown of FUBP1 in cells induced the formation of MDM2-ALT1, a stress-induced splice variant of MDM2, even under normal conditions. These results indicate that FUBP1 is also a strong positive splicing regulator that facilitates efficient splicing of the MDM2 pre-mRNA by binding its introns. These findings are the first report describing the regulation of alternative splicing of MDM2 mediated by the oncogenic factor FUBP1.
authors.affiliation: From the Center for Childhood Cancer, Research Institute at Nationwide Children's Hospital, Columbus, Ohio 43205 and the Department of Pediatrics, Molecular, Cellular and Developmental Biology Program, and Center for RNA Biology, Wexner Medical Center, The Ohio State University, Columbus, Ohio 43210.
authors.forename: Aishwarya G
authors.initials: AG
authors.lastname: Jacob
authors.affiliation: From the Center for Childhood Cancer, Research Institute at Nationwide Children's Hospital, Columbus, Ohio 43205 and the Department of Pediatrics, Molecular, Cellular and Developmental Biology Program, and.
authors.forename: Ravi K
authors.initials: RK
authors.lastname: Singh
authors.affiliation: From the Center for Childhood Cancer, Research Institute at Nationwide Children's Hospital, Columbus, Ohio 43205 and Center for RNA Biology, Wexner Medical Center, The Ohio State University, Columbus, Ohio 43210.
authors.forename: Fuad
authors.initials: F
authors.lastname: Mohammad
authors.affiliation: From the Center for Childhood Cancer, Research Institute at Nationwide Children's Hospital, Columbus, Ohio 43205 and the Department of Pediatrics, Molecular, Cellular and Developmental Biology Program, and.
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Bebee
authors.affiliation: From the Center for Childhood Cancer, Research Institute at Nationwide Children's Hospital, Columbus, Ohio 43205 and the Department of Pediatrics, Molecular, Cellular and Developmental Biology Program, and Center for RNA Biology, Wexner Medical Center, The Ohio State University, Columbus, Ohio 43210 Dawn.Chandler@nationwidechildrens.org.
authors.forename: Dawn S
authors.initials: DS
authors.lastname: Chandler
mesh_terms: D017398:Alternative Splicing / Q000502:physiology*; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007438:Introns / Q000502:physiology*; D009687:Nuclear Proteins / Q000096:biosynthesis* / Q000235:genetics; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins; D015534:Trans-Activators; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D004268:DNA-Binding Proteins; C492258:FUBP1 protein, human; C471297:MAML2 protein, human; C489427:MYC protein, human; D009687:Nuclear Proteins; D016271:Proto-Oncogene Proteins c-myc; D012322:RNA Precursors; D016601:RNA-Binding Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D004265:DNA Helicases
keywords: Cancer; MDM2; Oncogene; RNA; RNA Splicing; RNA-Protein Interaction; RNA-binding Protein; p53