REACTION CONDITIONS AND KINETICS OF SELF-CLEAVAGE OF A RIBOZYME DERIVED FROM NEUROSPORA VS RNA

被引:64
作者
COLLINS, RA
OLIVE, JE
机构
[1] Department of Molecular and Medical Genetics, University of Toronto, Toronto, Ontario M5S 1A8
关键词
D O I
10.1021/bi00062a009
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
We have investigated the self-cleavage reaction performed by a ribozyme that contains 164 nucleotides of Neurospora VS RNA. Self-cleavage requires a divalent cation, magnesium being more effective than manganese or calcium. Spermidine or monovalent cations stimulate the reaction but cannot replace magnesium. The temperature optimum is rather broad, around 40-50-degrees-C. Unlike some other ribozymes, VS self-cleavage is inhibited by even low concentrations of urea or formamide. The rate of cleavage is the same from pH 5.5 to 8.9, suggesting either that hydroxide is not directly involved in the cleavage chemistry or that a step that precedes the actual cleavage event is rate-limiting.
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收藏
页码:2795 / 2799
页数:5
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