Specific interference between two unrelated internal ribosome entry site elements impairs translation efficiency

Internal ribosome entry site (IRES) elements allow simultaneous synthesis of multiple proteins in eukaryotic cells. Here, two unrelated IRESs that perform efficiently in bicistronic constructs, the picornavirus foot-and-mouth disease virus (FMDV) and the cellular immunoglobulin heavy chain binding p...

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Bibliographic Details
Published in:FEBS letters 2005-12, Vol.579 (30), p.6803-6808
Main Authors: Reigadas, Sandrine, Pacheco, Almudena, Ramajo, Jorge, de Quinto, Sonia López, Martinez-Salas, Encarnacion
Format: Article
Language:English
Subjects:
Animals
BiP
CAT
Cell Line
chloramphenicol acetyl transferase
Cricetinae
Eukaryotic mRNA
firefly luciferase
FLuc
FMDV
Foot-and-mouth disease virus
Foot-and-Mouth Disease Virus - genetics
HCV
Heat-Shock Proteins - genetics
HeLa Cells
Hepacivirus - genetics
Hepatitis C virus
Humans
immunoglobulin heavy chain binding protein
Internal initiation of translation
internal ribosome entry site element
IRES
Molecular Chaperones - genetics
Peptide Chain Initiation, Translational - genetics
Peptide Chain Initiation, Translational - physiology
Picornavirus
Polycistronic vectors
Protein Biosynthesis
Protein synthesis
renilla luciferase
Ribosomes - metabolism
RLuc
RNA Caps - metabolism
RNA, Viral - genetics
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Summary:Internal ribosome entry site (IRES) elements allow simultaneous synthesis of multiple proteins in eukaryotic cells. Here, two unrelated IRESs that perform efficiently in bicistronic constructs, the picornavirus foot-and-mouth disease virus (FMDV) and the cellular immunoglobulin heavy chain binding protein (BiP) IRES, were used to generate a tricistronic vector. Functional analysis of the tricistronic RNA evidenced that the efficiency of protein synthesis under the control of BiP IRES was lower than that of the FMDV IRES, relative to the efficiency measured in bicistronic vectors. A specific competition between these elements was verified using two separate mono- or bicistronic constructs in vivo and in vitro. In contrast, no interference was detected with the hepatitis C virus (HCV) IRES. The interference effect of FMDV IRES was observed in cis and trans, in support of competition for common transacting factors different than those used in cap- and HCV-dependent initiation.
ISSN:0014-5793
1873-3468
DOI:10.1016/j.febslet.2005.11.015