The Activity of the Coat Protein Promoter of Chloris Striate Mosaic Virus Is Enhanced by Its Own and C1-C2 Gene Products

Abstract

DNA fragments from the bidirectional promoter region of the geminivirus chloris striate mosaic virus (CSMV) were cloned into the pUC18-based vector, pG1 producing transcriptional fusions with the β-glucuronidase (GUS) gene and nopaline synthase terminator sequence. The relative activity of each promoter construct was analyzed by a GUS expression assay of extracts from Zea mays (maize) Black Mexican Sweet protoplasts coelectroporated with the GUS reporter constructs and constructs in which individual CSMV open reading frames (ORFs) were placed under control of a cauliflower mosaic virus 35 S promoter. Weak promoter activity was observed for the promoter of the C1 and C2 ORFs (C1-C2 gene) and for the promoter of the V1 ORF. The activity of these promoters was unaffected by coelectroporation with the CSMV ORF constructs. Moderate activity was observed for the promoter of the V2 ORF (coat protein gene) which was enhanced by coelectroporation of the C1-C2 ORF construct. Sequences within the C1-C2 gene responsible for transactivation of the V2 ORF promoter were mapped close to the A site of a conserved NTP-binding sequence pattern within the C2 ORF. To a lesser extent activity for the promoter of the V2 ORF was enhanced by the V2 ORF construct providing evidence for positive autoregulation of the CSMV coat protein gene.