Sequence-specific RNA recognition by an RGG motif connects U1 and U2 snRNP for spliceosome assembly.
Sequence-specific RNA recognition by an RGG motif connects U1 and U2 snRNP for spliceosome assembly.
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DOI：
10.1073/pnas.2114092119
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发表时间：
2022-02-08
影响因子：
11.1
通讯作者：
Allain FH
中科院分区：
文献类型：
--
作者：
de Vries T;Martelly W;Campagne S;Sabath K;Sarnowski CP;Wong J;Leitner A;Jonas S;Sharma S;Allain FH
前信使RNA （pre-mRNA）剪接是基因表达的关键调控步骤。剪接反应由剪接体介导，剪接体是一种由五个小核核糖核蛋白（snRNPs）组成的动态复合体，它分多个步骤组装到每个内含子上。我们提出了人类U1和U2 snRNP之间重要的蛋白质- rna相互作用的详细结构分析和支持功能数据。我们的结构表明，U2 snRNP亚基的内在无序精氨酸-甘氨酸（RGG/RG）丰富基序与U1 snRNP形成rna序列特异性连接。本研究拓宽了RNA结合蛋白中非结构化RGG/富rg基序的功能范围，并为剪接体组装的早期步骤提供了分子基础，这可能有助于开发针对剪接缺陷引起的疾病的创新治疗策略。在哺乳动物中，U1和U2小核核糖核蛋白（snRNPs）在剪接的早期阶段相互作用的结构基础仍然是难以捉摸的。SF3A1的泛素样结构域（UBL）与U1 snRNP的茎环4 （U1- sl4）的结合有助于这种相互作用。在这里，我们确定了SF3A1的UBL和U1-SL4 RNA之间复合物的三维结构。我们的晶体学、核磁共振光谱和交联质谱数据表明，SF3A1-UBL特异性识别g1 - sl4的GCG/CGC RNA茎和顶端UUCG四环。体外和体内突变分析支持观察到的分子间接触，并证明SF3A1-UBL的羧基末端精氨酸-甘氨酸-甘氨酸-精氨酸（RGGR）基序通过插入RNA主槽特异性结合序列。因此，SF3A1-UBL/U1-SL4复合体的特征扩展了RNA结合域的库，并揭示了RGG/RG基序以序列特异性方式结合RNA的能力。
Pre-messenger RNA (pre-mRNA) splicing is a key regulatory step in gene expression. The splicing reaction is mediated by the spliceosome, a dynamic complex comprising five small nuclear ribonucleoproteins (snRNPs), which assembles onto each intron in multiple steps. We present detailed structural analysis and supporting functional data of an important protein–RNA interaction between human U1 and U2 snRNP. Our structure shows that an intrinsically disordered arginine-glycine (RGG/RG)–rich motif of a U2 snRNP subunit forms an RNA-sequence–specific connection with U1 snRNP. This study broadens the functional scope of unstructured RGG/RG-rich motifs in RNA binding proteins and provides a molecular basis of early steps of spliceosome assembly, which may help develop innovative therapeutic strategies against diseases originating from splicing defects. In mammals, the structural basis for the interaction between U1 and U2 small nuclear ribonucleoproteins (snRNPs) during the early steps of splicing is still elusive. The binding of the ubiquitin-like (UBL) domain of SF3A1 to the stem-loop 4 of U1 snRNP (U1-SL4) contributes to this interaction. Here, we determined the 3D structure of the complex between the UBL of SF3A1 and U1-SL4 RNA. Our crystallography, NMR spectroscopy, and cross-linking mass spectrometry data show that SF3A1-UBL recognizes, sequence specifically, the GCG/CGC RNA stem and the apical UUCG tetraloop of U1-SL4. In vitro and in vivo mutational analyses support the observed intermolecular contacts and demonstrate that the carboxyl-terminal arginine-glycine-glycine-arginine (RGGR) motif of SF3A1-UBL binds sequence specifically by inserting into the RNA major groove. Thus, the characterization of the SF3A1-UBL/U1-SL4 complex expands the repertoire of RNA binding domains and reveals the capacity of RGG/RG motifs to bind RNA in a sequence-specific manner.
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影响因子：
64.8
作者：
Pomeranz Krummel, Daniel A;Oubridge, Chris;Leung, Adelaine K W;Li, Jade;Nagai, Kiyoshi
通讯作者：
Nagai, Kiyoshi
影响因子：
16.6
作者：
Li X;Liu S;Jiang J;Zhang L;Espinosa S;Hill RC;Hansen KC;Zhou ZH;Zhao R
通讯作者：
Zhao R
影响因子：
16.6
作者：
Jutzi D;Campagne S;Schmidt R;Reber S;Mechtersheimer J;Gypas F;Schweingruber C;Colombo M;von Schroetter C;Loughlin FE;Devoy A;Hedlund E;Zavolan M;Allain FH;Ruepp MD
通讯作者：
Ruepp MD
影响因子：
56.9
作者：
CALNAN, BJ;TIDOR, B;FRANKEL, AD
通讯作者：
FRANKEL, AD
影响因子：
16
作者：
Hegele, Anna;Kamburov, Atanas;Stelzl, Ulrich
通讯作者：
Stelzl, Ulrich