Cap-independent translation initiation in Xenopus oocytes.
Eukaryotic cellular mRNAs contain a cap at their 5'-ends, but some viral and cellular mRNAs bypass the cap-dependent mechanism of translation initiation in favor of internal entry of ribosomes at specific RNA sequences. Cap-dependent initiation requires intact initiation factor eIF4G (formerly eIF-4gamma, eIF-4Fgamma or p220), whereas internal initiation can proceed with eIF4G cleaved by picornaviral 2A or L proteases. Injection of recombinant coxsackievirus B4 protease 2A into Xenopus oocytes led to complete cleavage of endogenous eIF4G, but protein synthesis decreased by only 35%. Co-injection of edeine reduced synthesis by >90%, indicating that eIF4G-independent synthesis involved ongoing initiation. The spectrum of endogenous proteins synthesized was very similar in the presence or absence of intact eIF4G. Translation of exogenous rabbit globin mRNA, by contrast, was drastically inhibited by eIF4G cleavage. The N-terminal cleavage product of eIF4G (cpN), which binds eIF4E, was completely degraded within 6-12 h, while the C-terminal cleavage product (cpC), which binds to eIF3 and eIF4A, was more stable over the same period. Thus, translation initiation of most endogenous mRNAs inXenopusoocytes requires no eIF4G, or perhaps only cpC, suggesting a cap-independent mechanism.
Keiper
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BD
Department of Biochemistry and Molecular Biology, Louisiana State University Medical Center, 1501 King's Highway, Shreveport, LA 71130-3932, USA.
Rhoads
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RE
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D003546:Cysteine Endopeptidases / Q000378:metabolism / Q000494:pharmacology; D004486:Edeine / Q000494:pharmacology; D005914:Globins / Q000235:genetics / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism*; D010448:Peptide Initiation Factors / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000235:genetics*; D012315:RNA Caps / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D011817:Rabbits; D014764:Viral Proteins; D014981:Xenopus
D010448:Peptide Initiation Factors; D012315:RNA Caps; D012333:RNA, Messenger; D014764:Viral Proteins; D004486:Edeine; D005914:Globins; D003546:Cysteine Endopeptidases; C057400:picornain 2A, Picornavirus