The Splicing Factor FUBP1 Is Required for the Efficient Splicing of Oncogene MDM2 Pre-mRNA

• Published in: The Journal of biological chemistry 2014-06, Vol.289 (25), p.17350-17364
• Main Authors: Jacob, Aishwarya G., Singh, Ravi K., Mohammad, Fuad, Bebee, Thomas W., Chandler, Dawn S.
• Format: Article
• Language: English
• Subjects: Alternative Splicing - physiology; Cancer; DNA Helicases - genetics; DNA Helicases - metabolism; DNA-Binding Proteins - biosynthesis; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; HeLa Cells; Humans; Introns - physiology; MDM2; Nuclear Proteins - biosynthesis; Nuclear Proteins - genetics; Oncogene; p53; Proto-Oncogene Proteins c-myc - genetics; Proto-Oncogene Proteins c-myc - metabolism; RNA; RNA Precursors - genetics; RNA Precursors - metabolism; RNA Splicing; RNA-binding Protein; RNA-Binding Proteins; RNA-Protein Interaction; Trans-Activators; Transcription Factors - biosynthesis; Transcription Factors - genetics
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M114.554717

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. Background:MDM2 is alternatively spliced into shorter isoforms under DNA damage and in several cancers through unknown mechanisms. Results: FUBP1 inactivation decreases splicing efficiency of an MDM2 minigene and causes exon skipping of endogenous MDM2 under normal conditions. Its overexpression attenuates damage-induced skipping of MDM2 exons. Conclusion: FUBP1 positively regulates efficient MDM2 splicing. Significance: This work uncovers an important mechanism regulating splicing efficiency of the oncogene MDM2.