Promoter analysis of the cell surface-abundant and hypoviral-regulated cryparin gene from Cryphonectria parasitica.
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 (-1257/-1158) and (-1107/-1008) with the characteristic AGGAGGA-N42-GAGAGGA and its inverted repeat TCCTCTC-N54-TCCTCCT, respectively, appeared to be specific binding sites for cellular factors.
Kim
Myoung-Ju
MJ
Institute for Molecular Biology and Genetics, Research Center of Bioactive Materials, Chonbuk National University, Jeonju 561-756, Korea.
Kwon
Bo-Ra
BR
Park
Seung-Moon
SM
Chung
Hea-Jong
HJ
Yang
Moon-Sik
MS
Churchill
Alice C L
AC
Van Alfen
Neal K
NK
Kim
Dae-Hyuk
DH
D001203:Ascomycota / Q000166:cytology / Q000235:genetics* / Q000821:virology*; D020029:Base Pairing; D001483:Base Sequence; D002462:Cell Membrane / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D005656:Fungal Proteins / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012328:RNA Viruses / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D014170:Transformation, Genetic
C074967:CRP protein, Cryphonectria parasitica; D005656:Fungal Proteins; D008565:Membrane Proteins; D012333:RNA, Messenger; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins