Suppression of PmRab11 inhibits YHV infection in Penaeus monodon

Fish & Shellfish Immunology
Akechai KongprajugChalermporn Ongvarrasopone

Abstract

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 5' 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 sig...Continue Reading

Datasets Mentioned

BETA
KY241479

Methods Mentioned

BETA
GTPase
PCR
PCRs
electrophoresis
X-ray

Software Mentioned

ImageJ
Expasy
Prenylation Prediction Suite
Gblocks
alignX
MUSCLE
PhyML
TreeDyn

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