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Molecular Genetics and Genomics

, Volume 287, Issue 11–12, pp 881–893 | Cite as

The 5′ untranslated region of the soybean cytosolic glutamine synthetase β1 gene contains prokaryotic translation initiation signals and acts as a translational enhancer in plants

  • Jose Luis Ortega
  • Olivia L. Wilson
  • Champa Sengupta-GopalanEmail author
Original Paper

Abstract

Glutamine synthetase (GS) catalyzes the synthesis of glutamine from glutamate and ammonia. In plants, it occurs as two major isoforms, a cytosolic form (GS1) and a nuclear encoded chloroplastic form. The focus of this paper is to determine the role of the 5′UTR of a GS1 gene. GS1 gene constructs with and without its 5′ and 3′ UTRs, driven by a constitutive promoter, were agroinfiltrated into tobacco leaves and the tissues were analyzed for both transgene transcript and protein accumulation. The constructs were also tested in an in vitro transcription/translation system and in Escherichia coli. Our results showed that while the 3′UTR functioned in the destabilization of the transcript, the 5′UTR acted as a translation enhancer in plant cells but not in the in vitro translation system. The 5′UTR of the GS1 gene when placed in front of a reporter gene (uidA), showed a 20-fold increase in the level of GUS expression in agroinfiltrated leaves when compared to the same gene construct without the 5′UTR. The 5′UTR-mediated translational enhancement is probably another step in the regulation of GS in plants. The presence of the GS1 5′UTR in front of the GS1 coding region allowed for its translation in E. coli suggesting the commonality of the translation initiation mechanism for this gene between plants and bacteria.

Keywords

Post-transcriptional regulation Translational control Glutamine synthetase β-Glucuronidase Agroinfiltration 

Notes

Acknowledgments

This work was supported by the National Institutes of Health (Grant number S06 GM08136-32), by the US Department of Agriculture (Grant number 2007-03596), and by the Agricultural Experimental Station at New Mexico State University. We thank Dr. Suman Bagga and Dr. Laura Rodriguez-Uribe for their helpful comments on the manuscript.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Jose Luis Ortega
    • 1
  • Olivia L. Wilson
    • 2
  • Champa Sengupta-Gopalan
    • 1
    Email author
  1. 1.Department of Plant and Environmental SciencesNew Mexico State UniversityLas CrucesUSA
  2. 2.Molecular Biology Graduate ProgramNew Mexico State UniversityLas CrucesUSA

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