Dutta Mandira, Su Yuan, Voth Gregory A, Stahelin Robert V
Department of Chemistry, Chicago Center for Theoretical Chemistry, Institute for Biophysical Dynamics, and James Frank Institute, The University of Chicago, Chicago, IL, USA.
Department of Medicinal Chemistry and Molecular Pharmacology and the Purdue Institute of Inflammation, Immunology and Infectious Disease, Purdue University, West Lafayette, IN, USA.
bioRxiv. 2023 Sep 15:2023.09.15.557899. doi: 10.1101/2023.09.15.557899.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a lipid-enveloped virus that acquires its lipid bilayer from the host cell it infects. SARS-CoV-2 can spread from cell to cell or from patient to patient by undergoing assembly and budding to form new virions. The assembly and budding of SARS-CoV-2 is mediated by several structural proteins known as envelope (E), membrane (M), nucleoprotein (N) and spike (S), which can form virus-like particles (VLPs) when co-expressed in mammalian cells. Assembly and budding of SARS-CoV-2 from the host ER-Golgi intermediate compartment is a critical step in the virus acquiring its lipid bilayer. To date, little information is available on how SARS-CoV-2 assembles and forms new viral particles from host membranes. In this study, we find the N protein can strongly associate with anionic lipids including phosphoinositides and phosphatidylserine. Moreover, lipid binding is shown to occur in the N protein C-terminal domain, which is supported by extensive analysis. Anionic lipid binding occurs for both the free and N oligomeric forms suggesting N can associate with membranes in the nucleocapsid form. Herein we present a lipid-dependent model based on , cellular and data for the recruitment of N to M assembly sites in the lifecycle of SARS-CoV-2.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）是一种包膜病毒，其脂质双层来自被感染的宿主细胞。SARS-CoV-2可通过组装和出芽形成新的病毒粒子，在细胞间或患者间传播。SARS-CoV-2的组装和出芽由几种结构蛋白介导，即包膜蛋白（E）、膜蛋白（M）、核蛋白（N）和刺突蛋白（S），当它们在哺乳动物细胞中共表达时可形成病毒样颗粒（VLP）。SARS-CoV-2从宿主内质网-高尔基体中间区室的组装和出芽是病毒获取脂质双层的关键步骤。迄今为止，关于SARS-CoV-2如何从宿主膜组装并形成新病毒颗粒的信息很少。在本研究中，我们发现N蛋白可与包括磷酸肌醇和磷脂酰丝氨酸在内的阴离子脂质强烈结合。此外，脂质结合发生在N蛋白的C末端结构域，这得到了广泛分析的支持。游离形式和N寡聚形式均发生阴离子脂质结合，表明N可以核衣壳形式与膜结合。在此，我们基于细胞和实验数据，提出了一种脂质依赖性模型，用于解释SARS-CoV-2生命周期中N蛋白被招募到M蛋白组装位点的过程。
