The 5' untranslated region of the VR-ACS1 mRNA acts as a strong translational enhancer in plants

Wever, Willem, McCallum, Emily J., Chakravorty, David, Cazzonelli, Christopher I. and Botella, José R. (2010) The 5' untranslated region of the VR-ACS1 mRNA acts as a strong translational enhancer in plants. Transgenic Research, 19 4: 667-674. doi:10.1007/s11248-009-9332-6


Author Wever, Willem
McCallum, Emily J.
Chakravorty, David
Cazzonelli, Christopher I.
Botella, José R.
Title The 5' untranslated region of the VR-ACS1 mRNA acts as a strong translational enhancer in plants
Formatted title
The 5′ untranslated region of the VR-ACS1 mRNA acts as a strong translational enhancer in plants
Journal name Transgenic Research   Check publisher's open access policy
ISSN 0962-8819
1573-9368
Publication date 2010-08-01
Year available 2009
Sub-type Article (original research)
DOI 10.1007/s11248-009-9332-6
Open Access Status Not yet assessed
Volume 19
Issue 4
Start page 667
End page 674
Total pages 8
Editor P. Christou
B. B. A. Whitelaw
Place of publication Dordrecht, Pays-Bas
Publisher Kluwer Academic
Language eng
Subject C1
970106 Expanding Knowledge in the Biological Sciences
060702 Plant Cell and Molecular Biology
Abstract The structure and function of untranslated mRNA leader sequences and their role in controlling gene expression remains poorly understood. Previous research has suggested that the 5' untranslated region (5'UTR) of the Vigna radiata aminocyclopropane-1-carboxylate synthase synthase (VR-ACS1) gene may function as a translational enhancer in plants. To test such hypothesis we compared the translation enhancing properties of three different 5'UTRs; those from the VR-ACS1, the chlorophyll a/b binding gene from petunia (Cab22L; a known translational enhancer) and the Vigna radiata pectinacetylesterase gene (PAE; used as control). Identical constructs in which the coding region of the beta-glucuronidase (GUS) gene was fused to each of the three 5'UTRs and placed under the control of the cauliflower mosaic virus 35S promoter were prepared. Transient expression assays in tobacco cell cultures and mung bean leaves showed that the VR-ACS1 and Cab22L 5'UTRs directed higher levels of GUS activity than the PAE 5'UTR. Analysis of transgenic Arabidopsis thaliana seedlings, as well as different tissues from mature plants, confirmed that while transcript levels were equivalent for all constructs, the 5'UTRs from the VR-ACS1 and Cab22L genes can increase GUS activity twofold to fivefold compared to the PAE 5'UTR, therefore confirming the translational enhancing properties of the VR-ACS1 5'UTR.
Formatted abstract
The structure and function of untranslated mRNA leader sequences and their role in controlling gene expression remains poorly understood. Previous research has suggested that the 5′ untranslated region (5′UTR) of the Vigna radiata aminocyclopropane-1-carboxylate synthase synthase (VR-ACS1) gene may function as a translational enhancer in plants. To test such hypothesis we compared the translation enhancing properties of three different 5′UTRs; those from the VR-ACS1, the chlorophyll a/b binding gene from petunia (Cab22L; a known translational enhancer) and the Vigna radiata pectinacetylesterase gene (PAE; used as control). Identical constructs in which the coding region of the β-glucuronidase (GUS) gene was fused to each of the three 5′UTRs and placed under the control of the cauliflower mosaic virus 35S promoter were prepared. Transient expression assays in tobacco cell cultures and mung bean leaves showed that the VR-ACS1 and Cab22L 5′UTRs directed higher levels of GUS activity than the PAE 5′UTR. Analysis of transgenic Arabidopsis thaliana seedlings, as well as different tissues from mature plants, confirmed that while transcript levels were equivalent for all constructs, the 5′UTRs from the VR-ACS1 and Cab22L genes can increase GUS activity twofold to fivefold compared to the PAE 5′UTR, therefore confirming the translational enhancing properties of the VR-ACS1 5′UTR.
© Springer Science+Business Media B.V. 2009
Keyword Translation efficiency
Promoter
Translational enhancer
Untranslated region
Mosaic virus-35S promoter
Arabidopsis-thaliana
Synthetic promoters
Gene-expression
High-efficiency
In-vivo
Protein
Tobacco
Sequence
Identification
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Published online: 9 October 2009

Document type: Journal Article
Sub-type: Article (original research)
Collections: 2010 Higher Education Research Data Collection
School of Biological Sciences Publications
 
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Created: Wed, 06 Jan 2010, 01:58:46 EST by Hayley Ware on behalf of School of Biological Sciences