Identification of RNA sequences and structures involved in site-specific cleavage of IGF-II mRNAs

被引:13
作者
Van Dijk, EL [1 ]
Sussenbach, JS [1 ]
Holthuizen, PE [1 ]
机构
[1] Univ Utrecht, Physiol Chem Lab, NL-3584 CG Utrecht, Netherlands
关键词
3 '-UTR; endonucleolytic cleavage; IGF-II; insulin-like growth factor-II; RNA-protein interactions; RNA secondary structure;
D O I
10.1017/S1355838298981316
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Insulin-like growth factor-ii (IGF-II) mRNAs are subject to site-specific endonucleolytic cleavage in the 3' untranslated region (UTR), rendering an unstable 5' cleavage product containing the coding region and a very stable 3' cleavage product of 1.8 kb consisting of the 3'-UTR sequence and the poly(A) tail. Previously, it was established that two widely separated elements in the 3'-UTR (elements I;and II), that can form a duplex structure, are necessary and sufficient for cleavage. To further investigate the sequence and secondary structure requirements for cleavage, we have introduced a number of mutations around the cleavage site and assayed their effects on cleavage. Several recognition determinants involved in the endonucleolytic cleavage of IGF-II mRNAs were identified. Mutational analysis around the cleavage site revealed that cleavage is sequence specific and that the cleavage site must be in a single-stranded conformation to allow efficient cleavage. In addition, we have identified an accessory protein that specifically interacts with a stem-loop structure located 133 to 73 nt upstream of the cleavage site.
引用
收藏
页码:1623 / 1635
页数:13
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