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首页> 外文期刊>Nucleic Acids Research >CUGBP2 directly interacts with U2 17S snRNP components and promotes U2 snRNA binding to cardiac troponin T pre-mRNA.
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CUGBP2 directly interacts with U2 17S snRNP components and promotes U2 snRNA binding to cardiac troponin T pre-mRNA.

机译：CUGBP2与U2 17S snRNP组件直接相互作用，并促进U2 snRNA与心脏肌钙蛋白T前mRNA结合。

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摘要

CUGBP2 (ETR-3/NAPOR/BRUNOL3) promotes inclusion of cardiac troponin T (cTNT) exon 5 via binding between positions 21 and 74 of the downstream intron. The molecular mechanism by which CUGBP2 activates cTNT exon 5 inclusion is unknown. Our results suggest that CUGBP2 promotes exon inclusion by a novel mechanism in which CUGBP2 directly interacts with components of the activated U2 snRNP and enhances binding of U2 snRNP to the branch site located upstream of the exon. Using an in vitro splicing assay, we show that recombinant CUGBP2 enhances complex A formation of a cTNT pre-mRNA. Enhanced complex A assembly requires both the upstream and downstream introns consistent with dual requirements for the downstream CUGBP2-binding site and an upstream branch site for U2 snRNP binding. We also show that CUGBP2 enhances binding of U2 snRNA to the cTNT pre-mRNA consistent with enhanced complex A assembly. Purification of CUGBP2-interacting proteins using tandem affinity purification leads to the demonstration that the core 17S U2 snRNP components, SF3b145 and SF3b49 bind directly to CUGBP2. We conclude that CUGBP2 activates exon inclusion by forming direct interactions with components of the 17S snRNP complex and recruits and/or stabilizes binding of U2 snRNP.
机译：CUGBP2（ETR-3 / NAPOR / BRUNOL3）通过下游内含子的21和74位之间的结合促进心肌肌钙蛋白T（cTNT）外显子5的包涵。 CUGBP2激活cTNT外显子5包涵体的分子机制尚不清楚。我们的结果表明CUGBP2通过一种新颖的机制促进外显子包涵，其中CUGBP2与活化的U2 snRNP的成分直接相互作用，并增强U2 snRNP与外显子上游分支位点的结合。使用体外剪接测定，我们显示重组CUGBP2增强cTNT前mRNA的复杂A形成。增强的复合物A装配需要上游和下游内含子，这与下游CUGBP2结合位点和U2 snRNP结合的上游分支位点的双重要求一致。我们还显示，CUGBP2增强U2 snRNA与cTNT pre-mRNA的结合，与增强的复合物A组装一致。使用串联亲和纯化纯化与CUGBP2相互作用的蛋白质，这证明了核心17S U2 snRNP组件SF3b145和SF3b49直接与CUGBP2结合。我们得出结论，CUGBP2通过与17S snRNP复合物的成分形成直接相互作用来激活外显子包涵，并募集和/或稳定U2 snRNP的结合。

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