Mapping the fMet-tRNAfMet binding site of initiation factor IF2

被引:82
作者
Guenneugues, M
Caserta, E
Brandi, L
Spurio, R
Meunier, S
Pon, CL
Boelens, R
Gualerzi, CO
机构
[1] Univ Utrecht, Bijvoet Ctr Biomol Res, NL-3584 CH Utrecht, Netherlands
[2] Univ Camerino, Dept Biol MCA, Genet Lab, I-62032 Camerino, MC, Italy
关键词
NMR spectroscopy; protein-RNA interaction; site-directed mutagenesis; translation initiation;
D O I
10.1093/emboj/19.19.5233
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
The interaction between fMet-tRNA(f)(Met) and Bacillus stearothermophilus translation initiation factor IF2 has been characterized. We demonstrate that essentially all thermodynamic determinants governing the stability and the specificity of this interaction are localized within the acceptor hexanucleotide fMet-3'ACCAAC of the initiator tRNA and a fairly small area at the surface of the beta-barrel structure of the 90-amino acid C-terminal domain of IF2 (IF2 C-2), A weak but specific interaction between IF2 C-2 and formyl-methionyl was also demonstrated. The surface of IF2 C-2 interacting with fMet.tRNA(f)(Met) has been mapped using two independent approaches, site-directed mutagenesis and NMR spectroscopy, which yielded consistent results. The binding site comprises C668 and G715 located in a groove accommodating the methionyl side-chain, R700, in the vicinity of the formyl group, Y701 and K702 close to the acyl bond between fMet and tRNA(f)(Met), and the surface lined with residues K702-S660, along which the acceptor arm of the initiator tRNA spans in the direction 3' to 5'.
引用
收藏
页码:5233 / 5240
页数:8
相关论文
共 26 条
  • [21] The C-terminal subdomain (IF2 C-2) contains the entire fMet-tRNA binding site of initiation factor IF2
    Spurio, R
    Brandi, L
    Caserta, E
    Pon, CL
    Gualerzi, CO
    Misselwitz, R
    Krafft, C
    Welfle, K
    Welfle, H
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (04) : 2447 - 2454
  • [22] SPURIO R, 1993, TRANSLATIONAL APPARATUS, P241
  • [23] Takahashi H, 2000, NAT STRUCT BIOL, V7, P220
  • [24] Late events of translation initiation in bacteria: a kinetic analysis
    Tomsic, J
    Vitali, LA
    Daviter, T
    Savelsbergh, A
    Spurio, R
    Striebeck, P
    Wintermeyer, W
    Rodnina, MV
    Gualerzi, CO
    [J]. EMBO JOURNAL, 2000, 19 (09) : 2127 - 2136
  • [25] Effect of the amino acid attached to Escherichia coli initiator tRNA on its affinity for the initiation factor IF2 and on the IF2 dependence of its binding to the ribosome
    Wu, XQ
    RajBhandary, UL
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (03) : 1891 - 1895
  • [26] Protein recognition by NMR
    Wüthrich, K
    [J]. NATURE STRUCTURAL BIOLOGY, 2000, 7 (03) : 188 - 189