鉴定泡沫病毒融合过程中的中间步骤。

Identification of an Intermediate Step in Foamy Virus Fusion.

机构信息

Department of Chemistry, Ludwig Maximilians-Universität München, Butenandtstraße 5-13, 81377 München, Germany.

Center for Nano Science (CENS), Ludwig Maximilians-Universität München, Butenandtstraße 5-13, 81377 München, Germany.

出版信息

Viruses. 2020 Dec 21;12(12):1472. doi: 10.3390/v12121472.

DOI:10.3390/v12121472

PMID:33371254

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7766700/

Abstract

Viral glycoprotein-mediated membrane fusion is an essential step for productive infection of host cells by enveloped viruses; however, due to its rarity and challenges in detection, little is known about the details of fusion events at the single particle level. Here, we have developed dual-color foamy viruses (FVs) composed of eGFP-tagged prototype FV (PFV) Gag and mCherry-tagged Env of either PFV or macaque simian FV (SFVmac) origin that have been optimized for detection of the fusion process. Using our recently developed tracking imaging correlation (TrIC) analysis, we were able to detect the fusion process for both PFV and SFVmac Env containing virions. PFV Env-mediated fusion was observed both at the plasma membrane as well as from endosomes, whereas SFVmac Env-mediated fusion was only observed from endosomes. PFV Env-mediated fusion was observed to happen more often and more rapidly than as for SFVmac Env. Strikingly, using the TrIC method, we detected a novel intermediate state where the envelope and capsids are still tethered but separated by up to 400 nm before final separation of Env and Gag occurred.

摘要

病毒糖蛋白介导的膜融合是包膜病毒感染宿主细胞的有效步骤；然而，由于其稀有性和检测挑战，我们对融合事件在单个颗粒水平的细节知之甚少。在这里，我们开发了双色泡沫病毒（FV），由 GFP 标记的原型 FV（PFV）Gag 和 mCherry 标记的 PFV 或猕猴猴猿猴泡沫病毒（SFVmac）来源的 Env 组成，这两种病毒经过优化，可用于检测融合过程。使用我们最近开发的跟踪成像相关（TrIC）分析，我们能够检测到含有 PFV 和 SFVmac Env 的病毒粒子的融合过程。PFV Env 介导的融合不仅发生在质膜上，也发生在内体中，而 SFVmac Env 介导的融合仅发生在内体中。PFV Env 介导的融合比 SFVmac Env 介导的融合发生得更频繁、更快。引人注目的是，使用 TrIC 方法，我们检测到了一种新的中间状态，在这种状态下，包膜和衣壳仍然被束缚，但在 Env 和 Gag 最终分离之前，它们之间的距离可以分离多达 400nm。