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剪接体相关蛋白SAP49的秀丽隐杆线虫同源物的多顺反子表达及RNA结合特异性

Polycistronic expression and RNA-binding specificity of the C. elegans homologue of the spliceosome-associated protein SAP49.

作者信息

Tanaka Y, Ohta A, Terashima K, Sakamoto H

机构信息

Department of Biology, Faculty of Science, Kobe University, Nada-ku.

出版信息

J Biochem. 1997 Apr;121(4):739-45. doi: 10.1093/oxfordjournals.jbchem.a021648.

Abstract

Splicing of mRNA precursors (pre-mRNAs) occurs in a multimolecular complex, termed spliceosome, which is comprised of pre-mRNA, small nuclear ribonucleoprotein particles (snRNPs), and other protein factors including spliceosome-associated proteins (SAPs). SAP49 is thought to be a subunit of the essential splicing factor SF3b and is involved in U2 snRNP function in mammalian cells. We have isolated a Caenorhabditis elegans cDNA encoding an RNA-binding protein with two RNA recognition motifs (RRMs) which shows extensive similarity to the human SAP49. The primary transcript for this C. elegans SAP49 homologue (cSAP49) seems to contain at least two additional cistrons and can be processed into three different mature mRNAs by trans-splicing. The cSAP49 mRNA, like other mRNAs in the same polycistronic unit, is expressed in most of the developmental stages, consistent with its putative essential function for mRNA splicing. By means of an in vitro RNA selection system, we demonstrate that cSAP49 possesses specific RNA-binding activity which resides in its second RRM.

摘要

mRNA前体(前体mRNA)的剪接发生在一种多分子复合物中,称为剪接体,它由前体mRNA、小核核糖核蛋白颗粒(snRNP)以及其他蛋白质因子组成,包括剪接体相关蛋白(SAP)。SAP49被认为是必需剪接因子SF3b的一个亚基,并参与哺乳动物细胞中的U2 snRNP功能。我们分离出了一种秀丽隐杆线虫cDNA,它编码一种具有两个RNA识别基序(RRM)的RNA结合蛋白,该蛋白与人类SAP49具有广泛的相似性。这种秀丽隐杆线虫SAP49同源物(cSAP49)的初级转录本似乎包含至少两个额外的顺反子,并且可以通过反式剪接加工成三种不同的成熟mRNA。cSAP49 mRNA与同一多顺反子单元中的其他mRNA一样,在大多数发育阶段都有表达,这与其对mRNA剪接的假定基本功能一致。通过体外RNA筛选系统,我们证明cSAP49具有特异性RNA结合活性,该活性存在于其第二个RRM中。

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