Alternative splicing of the neurofibromatosis type 1 pre-mRNA is regulated by the muscleblind-like proteins and the CUG-BP and ELAV-like factors.
Alternative splicing of the neurofibromatosis type 1 pre-mRNA is regulated by the muscleblind-like proteins and the CUG-BP and ELAV-like factors.
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DOI：
10.1186/1471-2199-13-35
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发表时间：
2012-12-10
影响因子：
--
通讯作者：
Lou H
中科院分区：
文献类型：
--
作者：
Fleming VA;Geng C;Ladd AN;Lou H
选择性剪接通常受到涉及许多蛋白质因子和顺式作用RNA序列元件的复杂调控。一个主要的挑战是确定所有的蛋白质球员，并确定他们如何控制一个特定的外显子的组合方式的替代表达。肌盲样蛋白（MBNL）和CUG-BP和ELAV样蛋白家族（CELF）是剪接调节蛋白，其在几个替代外显子的调节中起拮抗剂的作用。目前，已知只有有限数量的MBNL和CELF的共同靶标受这两组蛋白质的拮抗调节。最近，我们确定了1型神经纤维瘤病（NF 1）外显子23 a作为CELF蛋白负调控的新靶点。在这里，我们报告，MBNL家族成员是这个外显子的正调控。MBNL蛋白的过表达促进了低MBNL表达细胞系中外显子23 a的包含，并且高MBNL表达细胞系中MBNL 1和MBNL 2家族成员的同时siRNA介导的敲低促进了外显子23 a的跳跃。重要的是，这两组蛋白在调节外显子23 a的包含方面相互拮抗。此外，我们分析了这些蛋白质的结合位点的内含子序列上游的外显子23 a的UV交联测定。我们表明，在体外，除了先前确定的首选结合序列UGCUGU，MBNL蛋白需要的最佳结合的相邻序列。这项研究沿着我们先前的工作，证实了Hu、CELF、TIA-1和TIAR蛋白在NF 1外显子23 a调节中的作用，确定了该外显子处于严格、复杂的控制之下。
Alternative splicing is often subjected to complex regulatory control that involves many protein factors and cis-acting RNA sequence elements. One major challenge is to identify all of the protein players and define how they control alternative expression of a particular exon in a combinatorial manner. The Muscleblind-like (MBNL) and CUG-BP and ELAV-Like family (CELF) proteins are splicing regulatory proteins, which function as antagonists in the regulation of several alternative exons. Currently only a limited number of common targets of MBNL and CELF are known that are antagonistically regulated by these two groups of proteins. Recently, we identified neurofibromatosis type 1 (NF1) exon 23a as a novel target of negative regulation by CELF proteins. Here we report that MBNL family members are positive regulators of this exon. Overexpression of MBNL proteins promote exon 23a inclusion in a low MBNL-expressing cell line, and simultaneous siRNA-mediated knockdown of MBNL1 and MBNL2 family members in a high MBNL-expressing cell line promotes exon 23a skipping. Importantly, these two groups of proteins antagonize each other in regulating inclusion of exon 23a. Furthermore, we analyzed the binding sites of these proteins in the intronic sequences upstream of exon 23a by UV cross-linking assays. We show that in vitro, in addition to the previously identified preferred binding sequence UGCUGU, the MBNL proteins need the neighboring sequences for optimal binding. This study along with our previous work that demonstrated roles for Hu, CELF, and TIA-1 and TIAR proteins in the regulation of NF1 exon 23a establish that this exon is under tight, complex control.
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影响因子：
14.9
作者：
Barron, Victoria A.;Zhu, Hui;Lou, Hua
通讯作者：
Lou, Hua
影响因子：
2.5
作者：
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通讯作者：
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影响因子：
14.9
作者：
Kino, Yoshihiro;Washizu, Chika;Ishiura, Shoichi
通讯作者：
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影响因子：
1.2
作者：
Kanadia, RN;Urbinati, CR;Swanson, MS
通讯作者：
Swanson, MS
影响因子：
5.3
作者：
ANDERSEN, LB;BALLESTER, R;COLLINS, FS
通讯作者：
COLLINS, FS