Translation eukaryotic initiation factor 4G recognizes a specific structural element within the internal ribosome entry site of encephalomyocarditis virus RNA.
A complex of eukaryotic initiation factors (eIFs) 4A, 4E, and 4G (collectively termed eIF4F) plays a key role in recruiting mRNAs to ribosomes during translation initiation. The site of ribosomal entry onto most mRNAs is determined by interaction of the 5'-terminal cap with eIF4E; eIFs 4A and 4G may facilitate ribosomal entry by modifying mRNA structure near the cap and by interacting with ribosome-associated factors. eIF4G recruits uncapped encephalomyocarditis virus (EMCV) mRNA to ribosomes without the involvement of eIF4E by binding directly to the approximately 450-nucleotide long EMCV internal ribosome entry site (IRES). We have used chemical and enzymatic probing to map the eIF4G binding site to a structural element within the J-K domain of the EMCV IRES that consists of an oligo(A) loop at the junction of three helices. The oligo(A) loop itself is not sufficient to form stable complexes with eIF4G since alteration of its structural context abolished its interaction with eIF4G. Addition of wild type or trans-dominant mutant forms of eIF4A to binary IRES.eIF4G complexes did not further alter the pattern of chemical/enzymatic modification of the IRES.
Kolupaeva
V G
VG
A. N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, 119899 Moscow, Russia.
Pestova
T V
TV
Hellen
C U
CU
Shatsky
I N
IN
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D004686:Encephalomyocarditis virus / Q000235:genetics*; D039601:Eukaryotic Initiation Factor-4A; D039603:Eukaryotic Initiation Factor-4G; D039642:Eukaryotic Initiation Factors; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010448:Peptide Initiation Factors / Q000378:metabolism*; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D011817:Rabbits; D012270:Ribosomes / Q000378:metabolism*; D013329:Structure-Activity Relationship
D039603:Eukaryotic Initiation Factor-4G; D039642:Eukaryotic Initiation Factors; D010448:Peptide Initiation Factors; D012367:RNA, Viral; C031799:eIF-4B; D039601:Eukaryotic Initiation Factor-4A