Conversion of 48S translation preinitiation complexes into 80S initiation complexes as revealed by toeprinting

被引：59

作者：

Dmitriev, SE ^{[1
]}

Pisarev, AV ^{[1
]}

Rubtsova, MP ^{[1
]}

Dunaevsky, YE ^{[1
]}

Shatsky, IN ^{[1
]}

机构：

[1] Moscow MV Lomonosov State Univ, Belozersky Inst Physicochem Biol, Moscow 119899, Russia

来源：

FEBS LETTERS | 2003年 / 533卷 / 1-3期

基金：

俄罗斯基础研究基金会;

关键词：

translation initiation; AUG selection; toeprint assay; eukaryote;

D O I：

10.1016/S0014-5793(02)03776-6

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：

A method of analysis of translation initiation complexes by toeprinting has recently acquired a wide application to investigate molecular mechanisms of translation initiation in eukaryotes. So far, this very fruitful approach was used when researchers did not aim to discriminate between patterns of toeprints for 48S and 80S translation initiation complexes. Here, using cap-dependent and internal ribosomal entry site (IRES)-dependent mRNAs, we show that the toeprint patterns for 48S and 80S complexes are distinct whether the complexes are assembled in rabbit reticulocyte lysate or from fully purified individual components. This observation allowed us to demonstrate for the first time a delay in the conversion of the 48S complex into the 80S complex for beta-globin and encephalomyocarditis virus (EMCV) RNAs, and to assess the potential of some 80S antibiotics to block polypeptide elongation. Besides, additional selection of the authentic initiation codon among three consecutive AUGs that follow the EMCV IRES was revealed at steps subsequent to the location of the initiation codon by the 40S ribosomal subunit. (C) 2002 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

引用

页码：99 / 104

页数：6

共 25 条

[1]

ANTHONY DD, 1992, J BIOL CHEM, V267, P1554

[2] RNA - PROTEIN INTERACTIONS WITHIN THE INTERNAL TRANSLATION INITIATION REGION OF ENCEPHALOMYOCARDITIS VIRUS-RNA [J].

BOROVJAGIN, AV ;

EZROKHI, MV ;

ROSTAPSHOV, VM ;

UGAROVA, TY ;

BYSTROVA, TF ;

SHATSKY, IN .

NUCLEIC ACIDS RESEARCH, 1991, 19 (18) :4999-5005

[3] EFFICIENT INITIATION OF MAMMALIAN MESSENGER-RNA TRANSLATION AT A CUG CODON [J].

DASSO, MC ;

JACKSON, RJ .

NUCLEIC ACIDS RESEARCH, 1989, 17 (16) :6485-6497

[4] A COMPLEX RNA SEQUENCE DETERMINES THE INTERNAL INITIATION OF ENCEPHALOMYOCARDITIS VIRUS-RNA TRANSLATION [J].

EVSTAFIEVA, AG ;

UGAROVA, TY ;

CHERNOV, BK ;

SHATSKY, IN .

NUCLEIC ACIDS RESEARCH, 1991, 19 (03) :665-671

[5] STUDIES ON THE MODE OF ACTION OF HYGROMYCIN-B, AN INHIBITOR OF TRANSLOCATION IN EUKARYOTES [J].

GONZALEZ, A ;

JIMENEZ, A ;

VAZQUEZ, D ;

DAVIES, JE ;

SCHINDLER, D .

BIOCHIMICA ET BIOPHYSICA ACTA, 1978, 521 (02) :459-469

[6] RECOMBINANT IRON-REGULATORY FACTOR FUNCTIONS AS AN IRON-RESPONSIVE-ELEMENT-BINDING PROTEIN, A TRANSLATIONAL REPRESSOR AND AN ACONITASE - A FUNCTIONAL ASSAY FOR TRANSLATIONAL REPRESSION AND DIRECT DEMONSTRATION OF THE IRON SWITCH [J].

GRAY, NK ;

QUICK, S ;

GOOSSEN, B ;

CONSTABLE, A ;

HIRLING, H ;

KUHN, LC ;

HENTZE, MW .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1993, 218 (02) :657-667

[7]

GRIFO JA, 1983, J BIOL CHEM, V258, P5804

[8]

HARTZ D, 1988, METHOD ENZYMOL, V164, P419

[9]

Hershey JWB, 2000, COLD SPRING HARBOR M, V39, P33

[10] GTP hydrolysis controls stringent selection of the AUG start codon during translation initiation in Saccharomyces cerevisiae [J].

Huang, HK ;

Yoon, H ;

Hannig, EM ;

Donahue, TF .

GENES & DEVELOPMENT, 1997, 11 (18) :2396-2413

← 1 2 3 →