Genome organization and definition of the Penaeus monodon viral responsive protein 15 (PmVRP15) promoter. Issue 93 (October 2019)

• Record Type: Journal Article
• Authors: Jaree, Phattarunda; Kawai, Taro; Lo, Chu-Fang; Tassanakajon, Anchalee; Somboonwiwat, Kunlaya
• Is Part Of: Fish & shellfish immunology. Issue 93(2019)
• Page Start: 997
• Page End: 1006
• Publication Date: 2019-10
• Subjects: Fishes -- Immunology -- Periodicals; Shellfish -- Immunology -- Periodicals; Poissons -- Immunologie -- Périodiques; Crustacés -- Immunologie -- Périodiques; 571.9617
• DOI: 10.1016/j.fsi.2019.08.026
• Languages: English
• ISSNs: 1050-4648

Abstract

Abstract: The viral responsive protein 15 from the black tiger shrimp Penaeus monodon ( Pm VRP15) is a highly responsive gene upon white spot syndrome virus (WSSV) challenge. It is identified from hemocyte and important for WSSV trafficking and assembly. However, the knowledge of PmVRP15 gene regulation is limited. In the present study, the genome organization and 5′upstream promoter sequences of PmVRP15 gene were investigated. The PmVRP15 gene was found to contain 4 exons interrupted by 3 introns and the start codon was located in the exon 2. The transcription start site and TATA box were also determined from the 5′ upstream sequence. By using the narrow down experiment, the 5′ upstream promoter active region was determined to be at the nucleotide positions -525 to +612. Mutagenesis of the putative transcription factor (TF) binding sites revealed that the binding site of interferon regulatory factor (IRF) (-495/-479) was a repressor-binding site whereas those of the octamer transcription factor 1 (Oct-1) (-275/-268) and the nuclear factor of activated T-cells transcription factor (NFAT) (-228/-223) were activator-binding sites. This is the first report on the transcription factors that might play essential roles in modulating the PmVRP15 gene expression. Nevertheless, the underlying regulation mechanism of PmVRP15 gene expression needs further investigation. Highlights: The Pm VRP15 gene contained 4 exons interrupted by 3 introns that the start codon is located at exon 2. The 5′-flanking region of Pm VRP15 gene was identified by genome walking. The promoter active region of Pm VRP15 gene was characterized. IRF site was the repressor-binding site identified on Pm VRP15 promoter by mutagenesis. Oct-1 and NFAT sites were the activator-binding sites on Pm VRP15 promoter identified by mutagenesis.