Rab5, an early endosomal protein required for yellow head virus infection of Penaeus monodon

Suppression of PmRab11 inhibits YHV infection in Penaeus monodon

“…Free YHV particles were collected by ultracentrifugation (100,000Âg) for 1 h. Virus pellets were suspended with 150 mM NaCl and stored at À80 C until used. The viral nucleic acid was purified from the YHV stock using high pure viral nucleic acid kit (Roache Diagnostics, Germany) and subjected to RT-PCR to determine viral titer using primers YHV_F: 5ʹ-CAAGGACCACCTGGTACCGGTTAAGAC-3ʹ and YHV_R: 5ʹ-GCGGAAACGACTGACGGCTACATTCAC-3ʹ [11].…”

“…The partial sequence of PmRab11 served as template to design primers for 5ʹ and 3ʹ RACE. 5ʹ and 3ʹ RACE were performed as previously described [11]. Briefly, 5ʹ RACE method was performed by using 5Rab11_R1 specific primer (Table 1) to generate the firststrand cDNAs by Superscript III ® reverse transcriptase (Invitrogen).…”

“…Specifically, the transportation of YHV particles required PmRab5 which is a key protein in vesicles transport to early endosome; whereas, PmRab7 is involved in YHV transportation from early endosome to late endosome and lysosome. The silencing effects of PmRab5 or PmRab7 in YHV-infected P. monodon inhibited YHV expression, suggesting that PmRab5 and PmRab7 are involved in intracellular trafficking of YHV [11,12].…”

“…Recently, the mechanism of YHV transportation has been proposed. After YHV enters into the cell, it is transported via endosomal compartments [11,12] and released its genome into the cytoplasm for replication. Then, the nucleocapsid is synthesized and transported to the ER-Golgi compartment where the envelope is formed [2,13].…”

“…In the active form, Rab protein is associated with membranes by hydrophobic geranylgeranyl groups at the C-terminal and recruited the specific effector molecules such as sorting adaptors, tethering factors, kinases, phosphatases and motor proteins in vesicle transport process [18]. Our previous studies found that several Rab proteins are hijacked by YHV [11,12]. Specifically, the transportation of YHV particles required PmRab5 which is a key protein in vesicles transport to early endosome; whereas, PmRab7 is involved in YHV transportation from early endosome to late endosome and lysosome.…”