Domain IV of elongation factor G from Thermus thermophilus is strictly required for translocation

被引:30
作者
Martemyanov, KA [1 ]
Gudkov, AT [1 ]
机构
[1] Russian Acad Sci, Inst Prot Res, Pushchino 142292, Russia
基金
俄罗斯基础研究基金会;
关键词
ribosome; elongation factor G function; translocation; mutagenesis;
D O I
10.1016/S0014-5793(99)00635-3
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
Two truncated variants of elongation factor G from Thermus thermophilus with deletion of its domain IV have been constructed and the mutated genes were expressed in Escherichia coli, The truncated factors were produced in a soluble form and retained a high thermostability. It was demonstrated that mutated factors possessed (1) a reduced affinity to the ribosomes with an uncleavable GTP analog and (2) a specific ribosome-dependent GTPase activity. At the same time, in contrast to the mild-type elongation factor G, they were incapable to promote translocation, The conclusions are drawn that (I) domain IV is not involved in the GTPase activity of elongation factor G, (2) it contributes to the binding of elongation factor G with the ribosome and (3) is strictly required for translocation, These results suggest that domain IV might be directly involved in translocation and GTPase activity of the factor is not directly coupled with translocation, (C) 1999 Federation of European Biochemical Societies.
引用
收藏
页码:155 / 159
页数:5
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