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Expression of poliovirus 2A pro in mammalian cells: effects on translation
Author(s) -
Aldabe Rafael,
Feduchi Elena,
Novoa Isabel,
Carrasco Luis
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)01269-9
Subject(s) - vesicular stomatitis virus , poliovirus , microbiology and biotechnology , transfection , biology , protein biosynthesis , internal ribosome entry site , virology , endoplasmic reticulum , virus , vaccinia , recombinant virus , rna , translation (biology) , recombinant dna , messenger rna , biochemistry , gene
Poliovirus protease 2A pro has been efficiently expressed in HeLa and COS cells upon transfection with vector pTM1‐2A and infection with the recombinant vaccinia virus bearing the T7 RNA polymerase. The expressed poliovirus protease localizes to the cytoplasm of the transfected cells, both in the endoplasmic reticulum and in vesicles scattered in the cytoplasm. Cleavage of p220, a component of initiation factor eIF‐4F, selectively occurs from 5 h post‐infection in transfected cells infected with the recombinant virus. This cleavage correlates in time with the profound inhibition observed in the synthesis of vaccinia virus proteins. A similar blockade of vesicular stomatitis virus translation takes place upon 2A pro expression. Finally, the synthesis of poliovirus protein 2C from a recombinant vaccinia virus that expresses this protein under the EMC untranslated leader region is not affected by the synthesis of 2A pro . These findings lend support to the idea that translation of capped mRNAs requires the integrity of p220, while this requirement is not observed when translation of a mRNA bearing a picornavirus leader region is assayed.