Functional characterization of the EMCV IRES in plants

Urwin, Peter, Yi, Li, Martin, Helen, Atkinson, Howard and Gilmartin, Philip M. (2000) Functional characterization of the EMCV IRES in plants. The Plant Journal, 24 (5). pp. 583-589. ISSN 0960-7412

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Abstract

The translation of eukaryotic messenger RNA is typically dependent upon the presence of an mGpppN cap structure at the 5' end of the transcript. However, several animal viruses, including the Picorna viruses, have been shown to exhibit cap-independent translation through the presence of an internal ribosome entry site or IRES. This IRES-mediated cap-independent internal translation initiation has been exploited to generate bicistronic transcripts that function in animal cells. Recently IRES elements have also been identified in a small number of vertebrate, insect and yeast cellular messenger RNAs although no such sequences have been identified in endogenous plant genes and there are no reports of animal virus derived IRES activity in plant cells. Here we have constructed a bicistronic gene containing both green fluorescent protein and luciferase open-reading frames separated by the encephalomyocarditis IRES element under the control of the CaMV 35S promoter. Northern analysis reveals expression of the bicistronic transcript and in vivo imaging of GFP and luciferase activities demonstrates the functional presence of both proteins. Western blot analysis confirms the independent translation of both reporter proteins. These data suggest that insertion of the encephalomyocarditis virus (EMCV) IRES element between two open-reading frames of a plant bicistronic transcript can mediate translation of the second open-reading frame. This activity is more apparent in the leaves, than in the roots, of transgenic seedlings carrying the bicistronic reporter gene construct.

Item Type: Article
Uncontrolled Keywords: encephalomyocarditis virus (ecmv),function,internal ribosome entry site (ires),tobacco,translation
Faculty \ School: Faculty of Science > School of Biological Sciences
Related URLs:
Depositing User: Pure Connector
Date Deposited: 01 Nov 2013 14:12
Last Modified: 21 Apr 2020 22:06
URI: https://ueaeprints.uea.ac.uk/id/eprint/44155
DOI: 10.1046/j.1365-313X.2000.00904.x

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