ANALYSIS OF MESSENGER-RNA 3' END FORMATION BY MODIFICATION INTERFERENCE - THE ONLY MODIFICATIONS WHICH PREVENT PROCESSING LIE IN AAUAAA AND THE POLY(A) SITE
ANALYSIS OF MESSENGER-RNA 3' END FORMATION BY MODIFICATION INTERFERENCE - THE ONLY MODIFICATIONS WHICH PREVENT PROCESSING LIE IN AAUAAA AND THE POLY(A) SITE
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DOI：
10.1002/j.1460-2075.1987.tb02764.x
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发表时间：
1987-12-01
期刊：
影响因子：
11.4
通讯作者：
WICKENS, M
中科院分区：
文献类型：
--
作者：
CONWAY, L;WICKENS, M
描述了一种修饰干扰方法，其中使用化学修饰的转录本来鉴定任何以合成RNA为底物的反应所需的碱基。这种技术提供的信息类似于从分析一组完整的点突变中获得的信息。使用SV40晚期前mRNAs，我们确定AAUAAA序列中任何碱基的修饰都可以防止体外切割、多聚腺苷化和预切割复合体的形成。向其添加聚(A)的A的修饰防止了聚腺苷基化，但不干扰预切割复合体的形成。Poly(A)位点下游的单一修饰不会显著影响切割效率。由于Poly(A)位点下游区域是切割和复杂形成所必需的(Conway和Wkins，1985；Zarkower和Wkins，1987b)，我们推断该下游区域的关键特征要么是弥漫的，要么是多余的。
A modification interference method is described in which chemically modified transcripts are used to identify bases required for any reaction for which synthetic RNA is a substrate. This technique provides information analogous to that obtained from the analysis of a complete set of point mutants. Using SV40 late pre-mRNAs, we determine that modification of any base in the AAUAAA sequence prevents cleavage, polyadenylation and formation of pre-cleavage complexes in vitro. Modification of the A to which poly(A) is added prevents polyadenylation, but does not interfere with formation of the pre-cleavage complex. No single modification downstream of the poly(A) site significantly affects cleavage efficiency. Since the region downstream of the poly(A) site is required for cleavage and complex formation (Conway and Wickens, 1985; Zarkower and Wickens, 1987b), we infer that the critical features of this downstream region are either diffuse or redundant.