Article
- The EMBO Journal (2008) 27, 1609 - 1621
- doi:10.1038/emboj.2008.90
Published online: 8 May 2008
Subject Category:
Ribosomal position and contacts of mRNA in eukaryotic translation initiation complexes
Andrey V Pisarev1,a, Victoria G Kolupaeva1,ab, Marat M Yusupov3, Christopher UT Hellen1 and Tatyana V Pestova1,2
- Department of Microbiology and Immunology, SUNY Downstate Medical Center, Brooklyn, NY, USA
- AN Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia
- Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, France
Correspondence to:
Tatyana V Pestova, Department of Microbiology and Immunology, SUNY Downstate Medical Center, 450 Clarkson Avenue, Box 44, Brooklyn, NY 11203, USA. Tel.: +718 2216121; Fax: 718 2702656; E-mail: tatyana.pestova@downstate.edu
aThese authors contributed equally to this work
bPresent address: Department of Microbiology, New York University School of Medicine, New York, NY 10016, USA
Received 21 February 2008; Accepted 10 April 2008
Abstract
The position of mRNA on 40S ribosomal subunits in eukaryotic initiation complexes was determined by UV crosslinking using mRNAs containing uniquely positioned 4-thiouridines. Crosslinking of mRNA positions +11 to ribosomal protein (rp) rpS2(S5p) and rpS3(S3p), and +9–+11 and +8–+9 to h18 and h34 of 18S rRNA, respectively, indicated that mRNA enters the mRNA-binding channel through the same layers of rRNA and proteins as in prokaryotes. Upstream of the P-site, the proximity of positions -3/-4 to rpS5(S7p) and h23b, -6/-7 to rpS14(S11p), and -8–-11 to the 3'-terminus of 18S rRNA (mRNA/rRNA elements forming the bacterial Shine–Dalgarno duplex) also resembles elements of the bacterial mRNA path. In addition to these striking parallels, differences between mRNA paths included the proximity in eukaryotic initiation complexes of positions +7/+8 to the central region of h28, +4/+5 to rpS15(S19p), and -6 and -7/-10 to eukaryote-specific rpS26 and rpS28, respectively. Moreover, we previously determined that eukaryotic initiation factor2
(eIF2) contacts position -3, and now report that eIF3 interacts with positions -8–-17, forming an extension of the mRNA-binding channel that likely contributes to unique aspects of eukaryotic initiation.
Keywords:
- 4-thiouridine,
- eIF3,
- mRNA,
- ribosome,
- translation initiation
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