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Titolo:
Composition and arrangement of genes define the strength of IRES-driven translation in bicistronic mRNAs
Autore:
Hennecke, M; Kwissa, M; Metzger, K; Oumard, A; Kroger, A; Schirmbeck, R; Reimann, J; Hauser, H;
Indirizzi:
GBF German Res Ctr Biotechnol, Dept Gene Regulat & Differentiat, D-38124 Braunschweig, Germany GBF German Res Ctr Biotechnol Braunschweig Germany D-38124 weig, Germany Univ Ulm, Inst Med Mikrobiol, D-89069 Ulm, Germany Univ Ulm Ulm Germany D-89069 m, Inst Med Mikrobiol, D-89069 Ulm, Germany
Titolo Testata:
NUCLEIC ACIDS RESEARCH
fascicolo: 16, volume: 29, anno: 2001,
pagine: 3327 - 3334
SICI:
0305-1048(20010815)29:16<3327:CAAOGD>2.0.ZU;2-T
Fonte:
ISI
Lingua:
ENG
Soggetto:
INTERNAL RIBOSOME ENTRY; FACTOR MESSENGER-RNA; MAMMALIAN-CELLS; RETROVIRAL VECTORS; EXPRESSION VECTORS; SITE IRES; INITIATION; EUKARYOTES; PROTEIN; TRANSCRIPTION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
52
Recensione:
Indirizzi per estratti:
Indirizzo: Hauser, H GBF German Res Ctr Biotechnol, Dept Gene Regulat & Differentiat,Mascheroder Weg 1, D-38124 Braunschweig, Germany GBF German Res Ctr Biotechnol Mascheroder Weg 1 Braunschweig Germany D-38124
Citazione:
M. Hennecke et al., "Composition and arrangement of genes define the strength of IRES-driven translation in bicistronic mRNAs", NUCL ACID R, 29(16), 2001, pp. 3327-3334

Abstract

In addition to the cap-dependent mechanism, eukaryotic initiation of translation can occur by a cap-independent mechanism which directs ribosomes to defined start codons enabled by internal ribosome entry site (IRES) elements. IRES elements from poliovirus and encephalomyocarditis virus are often used to construct bi- or oligocistronic expression vectors to co-express various genes from one mRNA. We found that while cap-dependent translation initiation from bicistronic mRNAs remains comparable to monocistronic expression, internal initiation mediated by these viral IRESs is often very inefficient. Expression of bicistronic expression vectors containing the hepatitisB virus core antigen (HBcAg) together with various cytokines in the secondcistron of bicistronic mRNAs gave rise to very low levels of the tested cytokines. On the other hand, the HBcAg was well expressed when positioned inthe second cistron. This suggests that the arrangement of cistrons in a bicistronic setting is crucial for IRES-dependent translation of the second cistron. A systematic examination of expression of reporter cistrons from bicistronic mRNAs with respect to position was carried out. Using the dual luciferase assay system we show that the composition of reading frames on a bicistronic mRNA and the order in which they are arranged define the strength of IRES-dependent translation. Although the cellular environment and the nature of the IRES element influence translation strength the dominant determinant is the nature and the arrangement of cistrons on the mRNA.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 23/09/20 alle ore 14:49:47