MBNL1 and PTB cooperate to repress splicing of Tpm1 exon 3.
MBNL1 and PTB cooperate to repress splicing of Tpm1 exon 3.
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DOI：
10.1093/nar/gkt168
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发表时间：
2013-05
影响因子：
14.9
通讯作者：
Smith CW
中科院分区：
文献类型：
--
作者：
Gooding C;Edge C;Lorenz M;Coelho MB;Winters M;Kaminski CF;Cherny D;Eperon IC;Smith CW
大鼠α-原肌球蛋白（Tpm1）基因的外显子3在平滑肌细胞中被抑制，从而允许包含互斥的伴侣外显子2。影响外显子3剪接抑制的两种关键元件：多嘧啶束结合蛋白（PTB）的结合位点和由UGC或CUG基序组成的其他负调控元件。在这里，我们发现UGC簇与肌盲样蛋白（MBNL）结合，MBNL作为Tpm1外显子3的抑制因子。我们发现MBNL1的n端区域，包含其四个CCCH锌指结构域，足以介导抑制。MBNL1的同一区域可以与PTB发生直接的蛋白-蛋白相互作用，MBNL的RNA结合促进了这种相互作用，显然是通过诱导MBNL的构象变化来实现的。此外，单分子分析表明，mbnl结合位点增加了PTB与自身位点的结合。我们的数据表明，Tpm1的平滑肌剪接是由rna -蛋白复合物的变构组装介导的，该复合物最低限度地包含PTB、MBNL及其同源rna结合位点。
Exon 3 of the rat α-tropomyosin (Tpm1) gene is repressed in smooth muscle cells, allowing inclusion of the mutually exclusive partner exon 2. Two key types of elements affect repression of exon 3 splicing: binding sites for polypyrimidine tract-binding protein (PTB) and additional negative regulatory elements consisting of clusters of UGC or CUG motifs. Here, we show that the UGC clusters are bound by muscleblind-like proteins (MBNL), which act as repressors of Tpm1 exon 3. We show that the N-terminal region of MBNL1, containing its four CCCH zinc-finger domains, is sufficient to mediate repression. The same region of MBNL1 can make a direct protein-to-protein interaction with PTB, and RNA binding by MBNL promotes this interaction, apparently by inducing a conformational change in MBNL. Moreover, single molecule analysis showed that MBNL-binding sites increase the binding of PTB to its own sites. Our data suggest that the smooth muscle splicing of Tpm1 is mediated by allosteric assembly of an RNA–protein complex minimally comprising PTB, MBNL and their cognate RNA-binding sites.
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影响因子：
14.9
作者：
Kino, Yoshihiro;Washizu, Chika;Ishiura, Shoichi
通讯作者：
Ishiura, Shoichi
影响因子：
11.4
作者：
Cherny, Dmitry;Gooding, Clare;Eperon, Giles E.;Coelho, Miguel B.;Bagshaw, Clive R.;Smith, Christopher W. J.;Eperon, Ian C.
通讯作者：
Eperon, Ian C.
影响因子：
3.7
作者：
Lahmann, Ines;Fabienke, Manuela;Arnold, Hans-Henning
通讯作者：
Arnold, Hans-Henning
DOI：
10.1083/jcb.200105044
发表时间：
2001-11-26
期刊：
The Journal of cell biology
影响因子：
--
作者：
Hüttelmaier S;Illenberger S;Grosheva I;Rüdiger M;Singer RH;Jockusch BM
通讯作者：
Jockusch BM
影响因子：
16
作者：
Charlet-B, N;Logan, P;Cooper, TA
通讯作者：
Cooper, TA