Activation-Dependent TRAF3 Exon 8 Alternative Splicing Is Controlled by CELF2 and hnRNP C Binding to an Upstream Intronic Element

被引:17
作者
Schultz, Astrid-Solveig [1 ]
Preussner, Marco [1 ]
Bunse, Mario [2 ]
Karni, Rotem [3 ]
Heyd, Florian [1 ]
机构
[1] Free Univ Berlin, Inst Chem & Biochem, Berlin, Germany
[2] Max Delbruck Ctr Mol Med, Berlin, Germany
[3] Hebrew Univ Jerusalem, Hadassah Med Sch, Dept Biochem & Mol Biol, Inst Med Res Israel Canada, Jerusalem, Israel
关键词
RNA binding proteins; RNA splicing; RNA; PROTEINS; IDENTIFICATION; REGULATORS; EXPRESSION; SWITCH; RIBONUCLEOPROTEIN; TRANSCRIPTION; STABILITY; U2AF26;
D O I
10.1128/MCB.00488-16
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Cell-type-specific and inducible alternative splicing has a fundamental impact on regulating gene expression and cellular function in a variety of settings, including activation and differentiation. We have recently shown that activationinduced skipping of TRAF3 exon 8 activates noncanonical NF-kappa B signaling upon T cell stimulation, but the regulatory basis for this splicing event remains unknown. Here we identify cis-and trans-regulatory elements rendering this splicing switch activation dependent and cell type specific. The cis-acting element is located 340 to 440 nucleotides upstream of the regulated exon and acts in a distance-dependent manner, since altering the location reduces its activity. A small interfering RNA screen, followed by cross-link immunoprecipitation and mutational analyses, identified CELF2 and hnRNP C as trans-acting factors that directly bind the regulatory sequence and together mediate increased exon skipping in activated T cells. CELF2 expression levels correlate with TRAF3 exon skipping in several model systems, suggesting that CELF2 is the decisive factor, with hnRNP C being necessary but not sufficient. These data suggest an interplay between CELF2 and hnRNP C as the mechanistic basis for activation-dependent alternative splicing of TRAF3 exon 8 and additional exons and uncover an intronic splicing silencer whose full activity depends on the precise location more than 300 nucleotides upstream of the regulated exon.
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页数:14
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