PUROMYCIN REACTION FOR THE A-SITE-BOUND PEPTIDYL-TRANSFER RNA

被引：44

作者：

SEMENKOV, Y ^{[1
]}

SHAPKINA, T ^{[1
]}

MAKHNO, V ^{[1
]}

KIRILLOV, S ^{[1
]}

机构：

[1] BP KONSTANTINOV NUCL PHYS INST, GATCHINA 188350, USSR

来源：

FEBS LETTERS | 1992年 / 296卷 / 02期

关键词：

70S RIBOSOME; A-SITE; PUROMYCIN REACTION; VIOMYCIN; SPONTANEOUS TRANSLOCATION;

D O I：

10.1016/0014-5793(92)80380-Y

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：

AcPhe2-tRNA(Phe) synthesized in 70S ribosomes after consecutive binding of AcPhe-tRNA(Phe) at the P sites and EF-Tu-directed binding of Phe-tRNA(Phe) at the A sites is able to react quantitatively with puromycin in the absence of EF-G. A detailed study of the kinetics of the puromycin reaction, its comparison with that of spontaneous translocation, the use of antibiotic viomycin as an effective inhibitor of spontaneous translocation revealed that, besides spontaneous translocation, this peptidyl-tRNA could react with puromycin being located at the A site. This leads to the conclusion that the transpeptidation reaction per se triggers conformational changes in the ribosomal complex bringing the 3'-end of a newly synthesized peptidyl-tRNA nearer to the peptidyl-site of the peptidyltransferase center. This is detected functionally as the ability of such an A site bound peptidyl-tRNA to react with puromycin. This reaction is highly pronounced at elevated (25-degrees-C) temperature but can be hardly detected at 0-degrees-C.

引用

页码：207 / 210

页数：4

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