Global Promotion of Alternative Internal Exon Usage by mRNA 3' End Formation Factors
Department of Molecular, Cell and Cancer Biology
Medical Subject Headings
*Alternative Splicing; Base Sequence; Cleavage And Polyadenylation Specificity Factor; Exons; Genes, Reporter; Green Fluorescent Proteins; HEK293 Cells; Humans; Nuclear Proteins; Protein Binding; RNA Splice Sites; RNA, Messenger; RNA-Binding Proteins; Repressor Proteins; Ribonucleoprotein, U1 Small Nuclear; Ribonucleoproteins; Sequence Analysis, RNA
Genetics | Molecular Biology | Molecular Genetics
The mechanisms that regulate alternative precursor mRNA (pre-mRNA) splicing are largely unknown. Here, we perform an RNAi screen to identify factors required for alternative splicing regulation by RBFOX2, an RNA-binding protein that promotes either exon inclusion or exclusion. Unexpectedly, we find that two mRNA 3' end formation factors, cleavage and polyadenylation specificity factor (CPSF) and SYMPK, are RBFOX2 cofactors for both inclusion and exclusion of internal exons. RBFOX2 interacts with CPSF/SYMPK and recruits it to the pre-mRNA. RBFOX2 and CPSF/SYMPK then function together to regulate binding of the early intron recognition factors U2AF and U1 small nuclear ribonucleoprotein particle (snRNP). Genome-wide analysis reveals that CPSF also mediates alternative splicing of many internal exons in the absence of RBFOX2. Accordingly, we show that CPSF/SYMPK is also a cofactor of NOVA2 and heterologous nuclear ribonucleoprotein A1 (HNRNPA1), RNA-binding proteins that also regulate alternative splicing. Collectively, our results reveal an unanticipated role for mRNA 3' end formation factors in global promotion of alternative splicing.
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Citation: Mol Cell. 2015 Jun 4;58(5):819-31. doi: 10.1016/j.molcel.2015.03.016. Epub 2015 Apr 23. Link to article on publisher's site
Misra, Ashish; Ou, Jianhong; Zhu, Lihua Julie; and Green, Michael R., "Global Promotion of Alternative Internal Exon Usage by mRNA 3' End Formation Factors" (2015). Molecular, Cell and Cancer Biology Publications. 31.