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Title

Translation of Sindbis Virus 26 S mRNA Does Not Require Intact Eukariotic Initiation Factor 4G

AuthorsCastelló, Alfredo; Sanz, Miguel Ángel CSIC ORCID; Molina, Susana; Carrasco Llamas, Luis CSIC ORCID
KeywordsAlphavirus translation
Sindbis virus
eIF4G
Regulation of translation
Translation initiation factors
Issue Date28-Nov-2005
PublisherElsevier
CitationJournal of Molecular Biology Vol. 355, Issue 5, 3 February 2006, Pages 942-956
AbstractThe infection of baby hamster kidney (BHK) cells by Sindbis virus gives rise to a drastic inhibition of cellular translation, while under these conditions the synthesis of viral structural proteins directed by the subgenomic 26 S mRNA takes place efficiently. Here, the requirement for intact initiation factor eIF4G for the translation of this subgenomic mRNA has been examined. To this end, SV replicons that contain the protease of human immunodeficiency virus type 1 (HIV-1) or the poliovirus 2Apro replacing the sequences of SV glycoproteins have been constructed. BHK cells electroporated with the different RNAs synthesize protein C and the corresponding protease at late times. Notably, the proteolysis of eIF4G by both proteases has little effect on the translation of the 26 S mRNA. In addition, recombinant viable SVs were engineered that encode HIV-1 PR or poliovirus 2A protease under the control of a duplicated late promoter. Viral protein synthesis at late times of infection by the recombinant viruses is slightly affected in BHK cells that contain proteolysed eIF4G. The translatability of SV genomic 49 S mRNA was assayed in BHK cells infected with a recombinant virus that synthesizes luciferase and transfected with a replicon that expresses poliovirus 2Apro. Under conditions where eIF4G has been hydrolysed significantly the translation of genomic SV RNA was deeply inhibited. These findings indicate a different requirement for intact eIF4G in the translation of genomic and subgenomic SV mRNAs. Finally, the translation of the reporter gene that encodes green fluorescent protein, placed under the control of a second duplicate late promoter, is also resistant to the cleavage of eIF4G. In conclusion, despite the presence of a cap structure in the 5′ end of the subgenomic SV mRNA, intact eIF4G is not necessary for its translation
Publisher version (URL)http://dx.doi.org/10.1016/j.jmb.2005.11.024
URIhttp://hdl.handle.net/10261/5044
DOI10.1016/j.jmb.2005.11.024
ISSN0022-2836
E-ISSN1089-8638
Appears in Collections:(CBM) Artículos

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