[Minor secondary-structure variation in the 5'-untranslated region of the beta-globin mRNA changes the concentration requirements for eIF2].
Nucleotide sequence changes increasing the number of paired bases without producing stable secondary structure elements in the 5'-untranslated region (5'-UTR) of the beta-globin mRNA had a slight effect on its translation in rabbit reticulocyte lysate at its low concentration and dramatically decreased translation efficiency at a high concentration. The removal of paired regions restored translation. Addition of purified eIF2 to the lysate resulted in equal translation efficiencies of templates differing in structure of 5'-UTR. A similar effect was observed for p50, a major mRNP protein. Other mRNA-binding initiation factors, eIF4F and eIF3B, had no effect on the dependence of translation efficiency on mRNA concentration. Analysis of the assembly of the 48S initiation complex from its purified components showed that less eIF2 is required for translation initiation on the beta-globin mRNA than on its derivative containing minor secondary structure elements in 5'-UTR. According to a model proposed, eIF2 not only delivers Met-tRNA, but it also stabilizes the complex of the 40S ribosome subunit with 5'-UTR, which is of particular importance for translation initiation on templates with structured 5'-UTR.
Dmitriev
S E
SE
Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, 119899 Russia.
Terenin
I M
IM
Rubtsova
M P
MP
Shatskiĭ
I N
IN
D020121:5' Untranslated Regions; D000818:Animals; D001483:Base Sequence; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism*; D039621:Eukaryotic Initiation Factor-3 / Q000378:metabolism; D039562:Eukaryotic Initiation Factor-4F / Q000378:metabolism; D005914:Globins / Q000235:genetics*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012358:RNA, Transfer, Met / Q000378:metabolism; D011817:Rabbits; D013329:Structure-Activity Relationship
D020121:5' Untranslated Regions; D015852:Eukaryotic Initiation Factor-2; D039621:Eukaryotic Initiation Factor-3; D039562:Eukaryotic Initiation Factor-4F; D012333:RNA, Messenger; D012358:RNA, Transfer, Met; D005914:Globins