The Department of Molecular, Cell, and Developmental Biology, University of California Santa Barbara, Santa Barbara, CA, USA.
Molecular, Cellular, and Developmental Biology, University of California Santa Barbara, Santa Barbara, CA, USA.
Methods Mol Biol. 2024;2808:129-140. doi: 10.1007/978-1-0716-3870-5_10.
Many negative-sense single-stranded RNA viruses within the order Mononegavirales harm humans. A common feature shared among cells infected by these viruses is the formation of subcellular membraneless structures called biomolecular condensates, also known as inclusion bodies (IBs), that form through a process called liquid-liquid phase separation (LLPS). Like many other membraneless organelles, viral IBs enrich a specific subset of viral and host proteins involved in the formation of viral particles. Elucidation of the properties and regulation of these IBs as they mature throughout the viral replication process are important for our understanding of viral replication, which may also lead to the development of alternative antiviral treatments. The protocol outlined in this chapter aims to characterize the intrinsic properties of LLPS within the measles virus (MeV, a member of Mononegavirales) IBs by using an imaging approach that fluorescently tags an IB-associated host protein. This method uses common laboratory techniques and is generalizable to any host factors as well as other viral systems.
许多属于单负链 RNA 病毒目（Mononegavirales）的病毒会对人类造成危害。这些病毒感染的细胞共有的一个特征是形成无膜亚细胞结构，称为生物分子凝聚物，也称为包涵体（IBs），它们通过一种称为液-液相分离（LLPS）的过程形成。与许多其他无膜细胞器一样，病毒 IBs 富含参与病毒粒子形成的特定的病毒和宿主蛋白亚群。阐明这些 IBs 在病毒复制过程中成熟时的特性和调控对于我们理解病毒复制非常重要，这也可能导致替代抗病毒治疗方法的发展。本章概述的方案旨在通过荧光标记与 IB 相关的宿主蛋白的成像方法来表征麻疹病毒（MeV，属于单负链 RNA 病毒目）IB 内的 LLPS 的固有特性。该方法使用常见的实验室技术，并且可推广应用于任何宿主因子以及其他病毒系统。
