Promotion of exon 6 inclusion in HuD pre-mRNA by Hu protein family members.
Promotion of exon 6 inclusion in HuD pre-mRNA by Hu protein family members.
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DOI：
10.1093/nar/gkq028
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发表时间：
2010-06
影响因子：
14.9
通讯作者：
Lou H
中科院分区：
文献类型：
--
作者：
Wang H;Molfenter J;Zhu H;Lou H
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蛋白促进替代外显子的跳跃。在这里，我们报告的第一个例子，胡蛋白促进列入一个选择性外显子，外显子6的HuD前mRNA。序列比对分析表明，存在几个保守的AU丰富的序列上游和下游的这个选择性剪接外显子。我们产生了人HuD外显子6微型基因报告构建体，其中包括这些保守序列。Hu蛋白过表达导致来自该报告基因和内源性HuD的外显子6包含显著增加。使用截短的和突变的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的序列是重要的。Hu蛋白的RNAi敲除减少了外显子6的包含。体外剪接测定表明，Hu蛋白直接在剪接水平上促进HuD外显子6的包含。我们的研究表明，Hu蛋白可以作为剪接增强子，并扩大了Hu蛋白作为剪接调节子的功能作用。
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.
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影响因子：
11.4
作者：
Lal, A;Kawai, T;Gorospe, M
通讯作者：
Gorospe, M
影响因子：
5.3
作者：
Anderson, KD;Sengupta, J;Perrone-Bizzozero, NI
通讯作者：
Perrone-Bizzozero, NI
DOI：
10.1073/pnas.96.17.9885
发表时间：
1999-08-17
影响因子：
11.1
作者：
Akamatsu, W;Okano, HJ;Okano, H
通讯作者：
Okano, H
影响因子：
11.4
作者：
Dredge, BK;Stefani, G;Darnell, RB
通讯作者：
Darnell, RB
影响因子：
16
作者：
Katsanou, V;Papadaki, O;Kontoyiannis, DL
通讯作者：
Kontoyiannis, DL