Characterization of the domains of E-coli initiation factor IF2 responsible for recognition of the ribosome

被引:49
作者
Moreno, JMP [1 ]
Dyrskjotersen, L [1 ]
Kristensen, JE [1 ]
Mortensen, KK [1 ]
Sperling-Petersen, HU [1 ]
机构
[1] Aarhus Univ, Inst Mol & Struct Biol, Dept Biostruct Chem, DK-8000 Aarhus C, Denmark
关键词
translation initiation factor 2; IF2; IF1; IF3; in vitro ribosomal binding assay;
D O I
10.1016/S0014-5793(99)00858-3
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
We have studied the interactions between the ribosome and the domains of Escherichia coli translation initiation factor 2, using an in vitro ribosomal binding assay with wild-type forms, N- and C-terminaI truncated forms of IF2 as well as isolated structural domains, A deletion mutant of the factor consisting of the two N-terminal domains of Tn, binds to both 30S and 50S ribosomal subunits as well as to 70S ribosomes. Furthermore, a truncated form of IF2, lacking the two N-terminal domains, binds to 30S ribosomal subunits in the presence of IF1, In addition, this N-terminal deletion mutant IF2 possess a low but significant affinity for the 70S ribosome which is increased by addition of IF1, The isolated C-terminal domain of IF2 has no intrinsic affinity for the ribosome nor does the deletion of this domain from IF2 affect the ribosomal binding capability of IF2, We conclude that the N-terminus of IF2 is required for optimal interaction of the factor with both 30S and 50S ribosomal subunits, A structural model for the interaction of IF2 with the ribosome is presented. (C) 1999 Federation of European Biochemical Societies.
引用
收藏
页码:130 / 134
页数:5
相关论文
共 40 条
[1]   3-DIMENSIONAL STRUCTURE OF THE RIBOSOMAL TRANSLOCASE - ELONGATION-FACTOR-G FROM THERMUS-THERMOPHILUS [J].
AEVARSSON, A ;
BRAZHNIKOV, E ;
GARBER, M ;
ZHELTONOSOVA, J ;
CHIRGADZE, Y ;
AL-KARADAGHI, S ;
SVENSSON, LA ;
LILJAS, A .
EMBO JOURNAL, 1994, 13 (16) :3669-3677
[2]   Visualization of elongation factor G on the Escherichia coli 70S ribosome:: The mechanism of translocation [J].
Agrawal, RK ;
Penczek, P ;
Grassucci, RA ;
Frank, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (11) :6134-6138
[3]   DIRECT CROSS-LINKS BETWEEN INITIATION FACTOR-I, FACTOR-II, AND FACTOR-III AND RIBOSOMAL-PROTEINS PROMOTED BY 2-IMINOTHIOLANE [J].
BOILEAU, G ;
BUTLER, P ;
HERSHEY, JWB ;
TRAUT, RR .
BIOCHEMISTRY, 1983, 22 (13) :3162-3170
[4]  
BOLLEN A, 1975, J BIOL CHEM, V250, P4310
[5]   Initiation factors of protein biosynthesis in bacteria and their structural relationship to elongation and termination factors [J].
Brock, S ;
Szkaradkiewicz, K ;
Sprinzl, M .
MOLECULAR MICROBIOLOGY, 1998, 29 (02) :409-417
[6]   THE UNUSUAL TRANSLATIONAL INITIATION CODON AUU LIMITS THE EXPRESSION OF THE INFC (INITIATION-FACTOR IF3) GENE OF ESCHERICHIA-COLI [J].
BROMBACH, M ;
PON, CL .
MOLECULAR & GENERAL GENETICS, 1987, 208 (1-2) :94-100
[7]   THE PROTEIN-SYNTHESIS INITIATION FACTOR-II G-DOMAIN - STUDY OF A FUNCTIONALLY ACTIVE C-TERMINAL 65-KILODALTON FRAGMENT OF IF2 FROM ESCHERICHIA-COLI [J].
CENATIEMPO, Y ;
DEVILLE, F ;
DONDON, J ;
GRUNBERGMANAGO, M ;
SACERDOT, C ;
HERSHEY, JWB ;
HANSEN, HF ;
PETERSEN, HU ;
CLARK, BFC ;
KJELDGAARD, M ;
LACOUR, TFM ;
MORTENSEN, KK ;
NYBORG, J .
BIOCHEMISTRY, 1987, 26 (16) :5070-5076
[8]   Prokaryotic and eukaryotic translation factors [J].
Clark, BFC ;
GrunbergManago, M ;
Gupta, NK ;
Hershey, JWB ;
Hinnebusch, AG ;
Jackson, RJ ;
Maitra, U ;
Mathews, MB ;
Merrick, WC ;
Rhoads, RE ;
Sonenberg, N ;
Spremulli, LL ;
Trachsel, H ;
Voorma, HO .
BIOCHIMIE, 1996, 78 (11-12) :1119-1122
[9]  
ELISH ME, 1988, J GEN MICROBIOL, V134, P1355
[10]  
FAKUNDING JL, 1973, J BIOL CHEM, V248, P8555