title: PmVRP15, a novel viral responsive protein from the black tiger shrimp, Penaeus monodon, promoted white spot syndrome virus replication.
abstract: Suppression subtractive hybridization of Penaeus monodon hemocytes challenged with white spot syndrome virus (WSSV) has identified the viral responsive gene, PmVRP15, as the highest up-regulated gene ever reported in shrimps. Expression analysis by quantitative real time RT-PCR revealed 9410-fold up-regulated level at 48 h post WSSV injection. Tissue distribution analysis showed that PmVRP15 transcript was mainly expressed in the hemocytes of shrimp. The full-length cDNA of PmVRP15 transcript was obtained and showed no significant similarity to any known gene in the GenBank database. The predicted open reading frame of PmVRP15 encodes for a deduced 137 amino acid protein containing a putative transmembrane helix. Immunofluorescent localization of the PmVRP15 protein revealed it accumulated around the nuclear membrane in all three types of shrimp hemocytes and that the protein was highly up-regulated in WSSV-infected shrimps. Double-stranded RNA interference-mediated gene silencing of PmVRP15 in P. monodon significantly decreased WSSV propagation compared to the control shrimps (injected with GFP dsRNA). The significant decrease in cumulative mortality rate of WSSV-infected shrimp following PmVRP15 knockdown was observed. These results suggest that PmVRP15 is likely to be a nuclear membrane protein and that it acts as a part of WSSV propagation pathway.
authors.affiliation: Applied Analytical Chemistry Research Unit, Department of Chemistry, Faculty of Science, King Mongkut's Institute of Technology Ladkrabang, Bangkok, Thailand.
authors.forename: Tipachai
authors.initials: T
authors.lastname: Vatanavicharn
authors.affiliation: Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.
authors.forename: Adisak
authors.initials: A
authors.lastname: Prapavorarat
authors.affiliation: Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.
authors.forename: Phattarunda
authors.initials: P
authors.lastname: Jaree
authors.affiliation: Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.
authors.forename: Kunlaya
authors.initials: K
authors.lastname: Somboonwiwat
authors.affiliation: Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.
authors.forename: Anchalee
authors.initials: A
authors.lastname: Tassanakajon
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D006434:Hemocytes / Q000378:metabolism / Q000821:virology; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000235:genetics; D033561:Penaeidae / Q000235:genetics* / Q000821:virology*; D021381:Protein Transport; D017422:Sequence Analysis, DNA; D014779:Virus Replication; D046848:White spot syndrome virus 1 / Q000502:physiology*
chemical_list: D018076:DNA, Complementary