Deletion analysis of the promoter for the cucumber necrosis virus 0.9-kb subgenomic RNA

Abstract

Sequences comprising the core promoter for the cucumber necrosis virus (CNV) 0.9-kb subgenomic RNA have been determined using deletion analysis and site-directed mutagenesis. The deletion studies indicate that the promoter lies within a region located 20 nucleotides upstream and 6 nucleotides downstream and including the subgenomic start site. Sequences further upstream or downstream of the core promoter do not appear to strongly affect promoter activity or viral RNA accumulation. Results of site-directed mutagenesis studies indicate that nucleotides immediately surrounding the subgenomic start site regulate promoter activity. Comparison of sequences within the CNV promoter region with the corresponding region of other tombusviruses shows that the tombusvirus promoter shares a region of near complete identity in 14 of the 26 core promoter nucleotides. Little similarity exists between the CNV 0.9-kb subgenomic RNA promoter and the region surrounding the transcription initiation site for the CNV 2.1-kb subgenomic RNA. Likewise, limited similarity occurs with the 5' region of CNV genomic RNA. Sequences similar to the ICR2-like motifs found in the promoters of several alphavirus-like (supergroup III) plant and animal viruses are not apparent. This study represents the first analysis of a subgenomic promoter from a member of supergroup II of positive-strand RNA viruses. © 1995 Academic Press, Inc.