TY - JOUR
T1 - Suppression of PmRab11 inhibits YHV infection in Penaeus monodon
AU - Kongprajug, Akechai
AU - Panyim, Sakol
AU - Ongvarrasopone, Chalermporn
N1 - Funding Information:
We would like to thank Asst. Prof. Dr. Kusol Pootanakit for critically reading the manuscript, Prof. Dr. Paisarn Sithigorngul and Dr. Phattara-Orn Havanapan for providing anti-YHV-gp64 monoclonal antibody and anti-β-tubulin antibody, respectively and Ms. Chaweewan Chimawei, Mrs. Suparb Hongthong and Ms. Punnee Tongboonsong for technical assistance on culturing shrimp. We also would like to thank Shrimp Genetic Improvement Center in Surat Thani province, Thailand for providing shrimp samples. This work was supported by grants from Mahidol University under the National Research University initiative (NRU) and Thailand Research Fund (BRG5780006 to C.O, DBG5980011 to S.P. and IRG 5780009). A.K. is supported by the 60th Year Supreme Reign of His Majesty King Bhumibol Adulyadej.
Publisher Copyright:
© 2017 Elsevier Ltd
PY - 2017/7
Y1 - 2017/7
N2 - Yellow head virus (YHV) is one of the most serious pathogens that causes worldwide shrimp production loss. It enters the cells via clathrin-mediated endocytosis and utilizes small GTPase Rab proteins such as PmRab5 and PmRab7 for intracellular trafficking. In this study, molecular cloning and functional analysis of Rab11 during YHV infection were investigated. PmRab11 cDNA was cloned by Rapid amplification of cDNA ends (RACEs). It contained two forms of sizes 1200 and 1050 bp distinct at the 5ʹ UTR. The coding region of PmRab11 was 645 bp, encoding 214 amino acids. It also demonstrated the characteristics of Rab11 proteins containing five GTP-binding domains, five Rab family domains, four Rab subfamily domains and a prenylation site at the C-terminus. Suppression of PmRab11 using dsRNA-PmRab11 either before or after YHV-challenge resulted in significant inhibition of YHV levels in the hemocytes and viral release in the supernatant in both mRNA and protein levels. In addition, the silencing effect of PmRab11 in YHV-infected shrimps resulted in a delay in shrimp mortality for at least 2 days. Immunofluorescence study showed co-localization between PmRab11 and YHV at 24–72 h post YHV-challenge. In contrast, the co-localization signals were absence in the PmRab11 knockdown hemocytes and the YHV signals accumulated at the perinuclear region at 24 h post YHV-challenge. Then, accumulation of YHV was hardly observed after 48–72 h. These results suggested that PmRab11 is required for YHV infection in shrimp.
AB - Yellow head virus (YHV) is one of the most serious pathogens that causes worldwide shrimp production loss. It enters the cells via clathrin-mediated endocytosis and utilizes small GTPase Rab proteins such as PmRab5 and PmRab7 for intracellular trafficking. In this study, molecular cloning and functional analysis of Rab11 during YHV infection were investigated. PmRab11 cDNA was cloned by Rapid amplification of cDNA ends (RACEs). It contained two forms of sizes 1200 and 1050 bp distinct at the 5ʹ UTR. The coding region of PmRab11 was 645 bp, encoding 214 amino acids. It also demonstrated the characteristics of Rab11 proteins containing five GTP-binding domains, five Rab family domains, four Rab subfamily domains and a prenylation site at the C-terminus. Suppression of PmRab11 using dsRNA-PmRab11 either before or after YHV-challenge resulted in significant inhibition of YHV levels in the hemocytes and viral release in the supernatant in both mRNA and protein levels. In addition, the silencing effect of PmRab11 in YHV-infected shrimps resulted in a delay in shrimp mortality for at least 2 days. Immunofluorescence study showed co-localization between PmRab11 and YHV at 24–72 h post YHV-challenge. In contrast, the co-localization signals were absence in the PmRab11 knockdown hemocytes and the YHV signals accumulated at the perinuclear region at 24 h post YHV-challenge. Then, accumulation of YHV was hardly observed after 48–72 h. These results suggested that PmRab11 is required for YHV infection in shrimp.
KW - Black tiger shrimp
KW - Exocytosis
KW - Recycling endosome
KW - Viral budding
KW - Viral transportation
UR - http://www.scopus.com/inward/record.url?scp=85019638941&partnerID=8YFLogxK
U2 - 10.1016/j.fsi.2017.05.039
DO - 10.1016/j.fsi.2017.05.039
M3 - Article
C2 - 28527895
AN - SCOPUS:85019638941
SN - 1050-4648
VL - 66
SP - 433
EP - 444
JO - Fish and Shellfish Immunology
JF - Fish and Shellfish Immunology
ER -