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首页> 外文期刊>Nucleic Acids Research >Promotion of exon 6 inclusion in HuD pre-mRNA by Hu protein family members
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Promotion of exon 6 inclusion in HuD pre-mRNA by Hu protein family members

机译：Hu蛋白家族成员促进HuD pre-mRNA中外显子6包涵

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摘要

The Hu RNA-binding protein family consists of four members: HuR/A, HuB, HuC and HuD. HuR expression is widespread. The other three neuron-specific Hu proteins play an important role in neuronal differentiation through modulating multiple processes of RNA metabolism. In the splicing events examined previously, Hu proteins promote skipping of the alternative exons. Here, we report the first example where Hu proteins promote inclusion of an alternative exon, exon 6 of the HuD pre-mRNA. Sequence alignment analysis indicates the presence of several conserved AU-rich sequences both upstream and downstream to this alternatively spliced exon. We generated a human HuD exon 6 mini-gene reporter construct that includes these conserved sequences. Hu protein over-expression led to significantly increased exon 6 inclusion from this reporter and endogenous HuD. Studies using truncated and mutant HuD exon 6 reporters demonstrate that two AU-rich sequences located downstream of exon 6 are important. RNAi knockdown of Hu proteins decreased exon 6 inclusion. An in vitro splicing assay indicates that Hu proteins promote HuD exon 6 inclusion directly at the level of splicing. Our studies demonstrate that Hu proteins can function as splicing enhancers and expand the functional role of Hu proteins as splicing regulators.
机译：Hu RNA结合蛋白家族由四个成员组成：HuR / A，HuB，HuC和HuD。 HuR表达广泛。其他三种神经元特异性Hu蛋白通过调节RNA代谢的多个过程在神经元分化中发挥重要作用。在先前检查的剪接事件中，Hu蛋白促进其他外显子的跳跃。在这里，我们报告第一个例子，其中Hu蛋白促进HuD pre-mRNA的外显子外显子6的包涵。序列比对分析表明在该交替剪接外显子的上游和下游均存在几个保守的富含AU的序列。我们生成了包含这些保守序列的人类HuD外显子6微型基因报告基因构建体。 Hu蛋白的过表达导致该报道基因和内源性HuD显着增加了第6外显子的包涵度。使用截短和突变的HuD外显子6报告基因的研究表明，位于外显子6下游的两个富含AU的序列很重要。 Hu蛋白的RNAi敲低减少了外显子6包含。体外剪接分析表明，Hu蛋白直接在剪接水平上促进HuD外显子6的包涵。我们的研究表明，Hu蛋白可以充当剪接增强剂，并扩展Hu蛋白作为剪接调节剂的功能。

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