Promoter Analysis of the Cell Surface-abundant and Hypoviral-regulated Cryparin Gene from Cryphonectria parasitica

  • Kim, Myoung-Ju (Institute for Molecular Biology and Genetics, Research Center of Bioactive Materials, Chonbuk National University) ;
  • Kwon, Bo-Ra (Institute for Molecular Biology and Genetics, Research Center of Bioactive Materials, Chonbuk National University) ;
  • Park, Seung-Moon (Institute for Molecular Biology and Genetics, Research Center of Bioactive Materials, Chonbuk National University) ;
  • Chung, Hea-Jong (Institute for Molecular Biology and Genetics, Research Center of Bioactive Materials, Chonbuk National University) ;
  • Yang, Moon-Sik (Institute for Molecular Biology and Genetics, Research Center of Bioactive Materials, Chonbuk National University) ;
  • Churchill, Alice C.L. (Department of Plant Pathology, Cornell University) ;
  • Van Alfen, Neal K. (Department of Plant Pathology, University of California) ;
  • Kim, Dae-Hyuk (Institute for Molecular Biology and Genetics, Research Center of Bioactive Materials, Chonbuk National University)
  • Received : 2008.06.11
  • Accepted : 2008.08.29
  • Published : 2008.11.30

Abstract

Cryparin, encoded as a single copy gene (Crp) of the chestnut blight fungus Cryphonectria parasitica, is the most abundant protein produced by this fungus. However, its accumulation is decreased remarkably in C. parastica strains containing the double-stranded (ds) RNA virus Cryphonectria hypovirus 1. To characterize the transcriptional regulatory element(s) for strong expression and viral regulation, promoter analysis was conducted. Serial deletion of the Crp promoter region resulted in a step-wise decrease in promoter activity, indicating a localized distribution of genetic elements in the cryparin promoter. Promoter analysis indicated two positive and a repressive cis-acting elements. Among them, the promoter region between nt -1,282 and -907 appeared to be necessary for hypoviral-mediated down-regulation. An electrophoretic mobility shift assay (EMSA) on the corresponding promoter region (-1,282/-907) indicated two regions at (-1,257/-1,158) and (-1,107/-1,008) with the characteristic AGGAGGA-N42-GAGAGGA and its inverted repeat TCCTCTC-N54-TCCTCCT, respectively, appeared to be specific binding sites for cellular factors.

Keywords

cryparin;Cryphonectria parasitica;hypovirus;hypovirulence;promoter analysis

Acknowledgement

Supported by : Korea Science and Engineering Foundation (KOSEF)

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