Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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Identification of a Promoter Motif Involved in Curtovirus Sense-Gene Expression in Transgenic Arabidopsis

  • Hur, Jingyung (Department of Plant Cellular and Molecular Biology and Plant Biotechnology Center, The Ohio State University) ;
  • Choi, Eunseok (Department of Genetic Engineering, Sungkyunkwan University) ;
  • Buckley, Kenneth J. (Plant Biotechnology Center, The Ohio State University) ;
  • Lee, Sukchan (Department of Genetic Engineering, Sungkyunkwan University) ;
  • Davis, Keith R. (James Graham Brown Cancer Center and Department of Pharmacology and Toxicology, University of Louisville School of Medicine)
  • Received : 2007.10.23
  • Accepted : 2008.03.17
  • Published : 2008.08.31
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Abstract

Expression of the seven open reading frames (ORFs) of single-stranded DNA Curtoviruses such as Beet curly top virus (BCTV) and Beet severe curly top virus (BSCTV) is driven by a bi-directional promoter. To investigate this bidirectional promoter activity with respect to viral late gene expression, transgenic Arabidopsis plants expressing a GUS reporter gene under the control of either the BCTV or BSCTV bi-directional promoter were constructed. Transgenic plants harboring constructs showed higher expression levels when the promoter of the less virulent BCTV was used than when the promoter of the more virulent BSCTV was used. In transgenic seedlings, the reporter gene constructs were expressed primarily in actively dividing tissues such as root tips and apical meristems. As the transgenic plants matured, reporter gene expression diminished but viral infection of mature transgenic plants restored reporter gene expression, particularly in transgenic plants containing BCTV virion-sense gene promoter constructs. A 30 base pair conserved late element (CLE) motif was identified that was present three times in tandem in the BCTV promoter and once in that of BSCTV. Progressive deletion of these repeats from the BCTV promoter resulted in decreased reporter gene expression, but BSCTV promoters in which one or two extra copies of this motif were inserted did not exhibit increased late gene promoter activity. These results demonstrate that Curtovirus late gene expression by virion-sense promoters depends on the developmental stage of the host plant as well as on the number of CLE motifs present in the promoter.

Keywords

Acknowledgement

Supported by : National Institutes of Health

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