Nucleic Acids Research, Vol 25, Issue 2 395-402, Copyright © 1997 by Oxford University Press
BD Keiper and RE Rhoads
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.
ARTICLES
Cap-independent translation initiation in Xenopus oocytes
Department of Biochemistry and Molecular Biology, Louisiana State University Medical Center, 1501 King's Highway, Shreveport, LA 71130- 3932, USA.
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