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Snap25 Regulates The Release Of The Rabies Virus In Nerve Cells Via Snare Complex-Mediated Membrane Fusion

文献类型： 外文期刊

作者： Yin, K; Li, YM; Ma, ZP; Yang, Y; Zhao, HZ; Liu, CY; Jin, M; Wudong, G; Sun, YM; Hang, TY; Zhang, H; Wang, FX; Wen, YJ

作者机构：

关键词： Rabies Virus; Glycoprotein; Snap25; Nerve Cells; Membrane Fusion

期刊名称： VETERINARY MICROBIOLOGY

ISSN： 0378-1135

年卷期： 2020 年 245 卷 0 期

页码：

收录情况： JCR（2021版）

摘要： Recent studies have reported that host proteins regulate Rabies virus (RABV) infection via distinct mechanisms. The abnormal neural function caused by RABV infection is related to the abnormal synaptic signal transmission in which the RABV glycoprotein (G) is involved. In the present study, two recombinant Rabies viruses (rRABVs), namely rSAD-SAD-Flag-G and rSAD-CVS-Flag-G, were established and rescued based on rSAD and verified by indirect fluorescence assay (IFA), and western blotting (WB). To investigate how the G protein interacts with synaptosomal-associated protein 25 (SNAP25), primary neuronal cells (PNC) of embryonic mice were cultured and infected with rRABVs. Immunoprecipitation (IP) and LC-MS/MS analysis of glycoprotein-binding proteins, which were flag tagged, were carried out to determine the interaction of G protein and soluble N-ethylmaleimide-sensitive factor attachment protein receptor proteins (SNARE) complex in PNC. G protein and the SNARE member SNAP25 were co-expressed in HEK293 cells or primary neuronal cells to investigate their colocalization. Knockdown of SNAP25 with small interfering RNA (siRNA) was conducted on mNA cells, and rRABV replication was observed by IFA, qRT-PCR, and virus titration. The results indicated that rRABVs were successfully rescued and grew well in PNC. Flag-tag IP and confocal microscopy demonstrated that SNAP25 works together with G protein and colocalizes with G on the cytomembrane of HEK293 cells. The downregulation of SNAP25, using RNA interference, resulted in a significant decrease in the number of viral mRNAs, viral proteins, and virus particles. Furthermore, the regression of SNAP25 did not affect the initial infection of the virus but reduced the infectivity of progeny virions.

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