The 5"² untranslated region of the VR-ACS1 mRNA acts as a strong translational enhancer in plants

Willem Wever; Emily J. McCallum; David Chakravorty; Christopher I. Cazzonelli; José R. Botella

doi:10.1007/s11248-009-9332-6

The 5"² untranslated region of the VR-ACS1 mRNA acts as a strong translational enhancer in plants

Willem Wever, Emily J. McCallum, David Chakravorty, Christopher I. Cazzonelli, José R. Botella

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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"² untranslated region (5"²UTR) 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"²UTRs; 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 Î²-glucuronidase (GUS) gene was fused to each of the three 5"²UTRs 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"²UTRs directed higher levels of GUS activity than the PAE 5"²UTR. 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"²UTRs from the VR-ACS1 and Cab22L genes can increase GUS activity twofold to fivefold compared to the PAE 5"²UTR, therefore confirming the translational enhancing properties of the VR-ACS1 5"²UTR.

Original language	English
Pages (from-to)	667-674
Number of pages	8
Journal	Transgenic Research
Volume	19
Issue number	4
DOIs	https://doi.org/10.1007/s11248-009-9332-6
Publication status	Published - 2010

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title = "The 5{"}² untranslated region of the VR-ACS1 mRNA acts as a strong translational enhancer in plants",

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{"}² untranslated region (5{"}²UTR) 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{"}²UTRs; 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 {\^I}²-glucuronidase (GUS) gene was fused to each of the three 5{"}²UTRs 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{"}²UTRs directed higher levels of GUS activity than the PAE 5{"}²UTR. 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{"}²UTRs from the VR-ACS1 and Cab22L genes can increase GUS activity twofold to fivefold compared to the PAE 5{"}²UTR, therefore confirming the translational enhancing properties of the VR-ACS1 5{"}²UTR.",

author = "Willem Wever and McCallum, {Emily J.} and David Chakravorty and Cazzonelli, {Christopher I.} and Botella, {Jos{\'e} R.}",

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journal = "Transgenic Research",

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T1 - The 5"² untranslated region of the VR-ACS1 mRNA acts as a strong translational enhancer in plants

AU - Wever, Willem

AU - McCallum, Emily J.

AU - Chakravorty, David

AU - Cazzonelli, Christopher I.

AU - Botella, José R.

PY - 2010

Y1 - 2010

N2 - 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"² untranslated region (5"²UTR) 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"²UTRs; 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 Î²-glucuronidase (GUS) gene was fused to each of the three 5"²UTRs 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"²UTRs directed higher levels of GUS activity than the PAE 5"²UTR. 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"²UTRs from the VR-ACS1 and Cab22L genes can increase GUS activity twofold to fivefold compared to the PAE 5"²UTR, therefore confirming the translational enhancing properties of the VR-ACS1 5"²UTR.

AB - 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"² untranslated region (5"²UTR) 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"²UTRs; 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 Î²-glucuronidase (GUS) gene was fused to each of the three 5"²UTRs 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"²UTRs directed higher levels of GUS activity than the PAE 5"²UTR. 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"²UTRs from the VR-ACS1 and Cab22L genes can increase GUS activity twofold to fivefold compared to the PAE 5"²UTR, therefore confirming the translational enhancing properties of the VR-ACS1 5"²UTR.

UR - http://handle.uws.edu.au:8081/1959.7/540802

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DO - 10.1007/s11248-009-9332-6

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JF - Transgenic Research

IS - 4

ER -

The 5"² untranslated region of the VR-ACS1 mRNA acts as a strong translational enhancer in plants

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