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Virol Res J 2017 Volume 1 Issue 3

International Virology Conference

October 30-31, 2017 | Toronto, Canada

Characterization of the entry mechanism of a novel protein transduction domain originated from

Betanodavirus

Junfeng Xie, Guohua Zhu, Jing Zhang and Runqing Huang

Sun Yat-Sen University, China

etanodavirus, also called nervous necrosis virus (NNV), is

the causative agent of a fish disease viral nervous necrosis.

VNN causes high mortality in marine fish at larvae and juvenile

stages resulting in heavy financial losses of marine aquaculture

industry. Base on the sequence analysis and 3D structure

elucidation of orange-spotted grouper nervous necrosis virus

(OGNNV), we found a peptide with the capsid protein can carry

foreign protein to enter fish cells. Through cell entry experiment

with different sections of the peptide fused with GFP, we

characterized a novel 14-aa peptide as protein transduction

domain (PTD) and named NNV-PTD. NNV-PTD can be fused

with foreign protein at N- or C-terminal without disturbing

the entry efficiency. In addition, NNV-PTD can enter different

types of fish cells, insect cells and mammalian cells as well as

enters fish cells with higher efficiency than well-known PTD

such as TAT, Penetratin and R8. NNV-PTD entered fish cells to

perinuclear regions within 4 h and can be blocked by culturing

at 4, indicating the endocytosis-dependent entry mechanism.

Therefore, we used the biochemical inhibitors blocking clathrin-

mediated endocytosis (CME), micropinocytosis, caveolae-

dependent endocytosis, cellular cholesterol, low-pH balance

and cytoskeleton to treat cells before entry assay. Finally, we

demonstrated thatNNV-PTDenters fishcells viaCMEdepending

on dynamin and macropinocytosis depending on myosin

and also the entry is cholesterol-, low-pH, and cytoskeleton-

denpendent at the intracellular traffic level. Together, this

work not only characterizes a novel high efficient PTD but also

identifies its entry mechanism, providing basic information for

further application of NNV-PTD in aquaculture.

Speaker Biography

Junfeng Xie has completed his PhD from Sun Yat-sen University, China. He is the

Associate Professor of School of Lifesciences, Sun Yat-sen University, USA. He is

focusing on the basic virology research of Betanodavirus and on the application study

of antiviral agents.