Dependence of the adenovirus tripartite leader on the p220 subunit of eukaryotic initiation factor 4F during in vitro translation. Effect of p220 cleavage by foot-and-mouth-disease-virus L-protease on in vitro translation.
The adenovirus tripartite leader (TPT) 5' untranslated region (5'UTR) allows translation in poliovirus-infected cells, in which the p220 subunit of eukaryotic initiation factor 4F is degraded. This p220-independent translation was investigated by measuring in vitro translation in a reticulocyte lysate of a reporter gene, chloramphenicol acetyltransferase, coupled to the TPT 5'UTR. The p220 subunit was degraded by translation of a foot-and-mouth-disease L-protease construct. Surprisingly, the TPT 5'UTR was dependent on intact p220, as are other naturally capped mRNA species. Translation of encephalomyocarditis virus RNA was p220 independent, as expected from its ability to support internal, cap-independent initiation. In vitro protein-synthesis experiments with purified initiation factors confirmed the dependence of TPT mRNA translation on eukaryotic initiation factor 4F. The relationship between adenovirus TPT-5'UTR-directed translation and poliovirus-induced host cell shut-off is discussed.
Thomas
A A
AA
Department of Molecular Cell Biology, University of Utrecht, The Netherlands.
Scheper
G C
GC
Kleijn
M
M
De Boer
M
M
Voorma
H O
HO
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D005537:Aphthovirus / Q000201:enzymology; D002474:Cell-Free System; D010450:Endopeptidases / Q000378:metabolism; D039562:Eukaryotic Initiation Factor-4F; D015967:Gene Expression Regulation, Viral; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D010442:Peptide Chain Initiation, Translational; D010448:Peptide Initiation Factors / Q000378:metabolism* / Q000648:ultrastructure; D012315:RNA Caps; D012367:RNA, Viral / Q000235:genetics*; D011817:Rabbits
D039562:Eukaryotic Initiation Factor-4F; D046911:Macromolecular Substances; D010448:Peptide Initiation Factors; D012315:RNA Caps; D012367:RNA, Viral; D010450:Endopeptidases