Cytoplasmic domain and enzymatic activity of ACE2 are not required for PI4KB dependent endocytosis entry of SARS-CoV-2 into host cells

Hang Yang; Huijun Yuan; Xiaohui Zhao; Meng Xun; Shangrui Guo; Nan Wang; Bing Liu; Hongliang Wang

doi:10.1016/j.virs.2022.03.003

June 2022

Hang Yang, Huijun Yuan, Xiaohui Zhao, Meng Xun, Shangrui Guo, Nan Wang, Bing Liu and Hongliang Wang. Cytoplasmic domain and enzymatic activity of ACE2 are not required for PI4KB dependent endocytosis entry of SARS-CoV-2 into host cells[J]. Virologica Sinica, 2022, 37(3): 380-389. doi: 10.1016/j.virs.2022.03.003

Citation: Hang Yang, Huijun Yuan, Xiaohui Zhao, Meng Xun, Shangrui Guo, Nan Wang, Bing Liu, Hongliang Wang. Cytoplasmic domain and enzymatic activity of ACE2 are not required for PI4KB dependent endocytosis entry of SARS-CoV-2 into host cells .VIROLOGICA SINICA, 2022, 37(3) : 380-389. http://dx.doi.org/10.1016/j.virs.2022.03.003

SARS-CoV-2病毒通过内吞途径感染宿主细胞依赖于PI4KB活性但不依赖于ACE2胞内区及酶催化活性位点

杨行 ^a ,
苑慧珺 ^a ,
赵小惠 ^a ,
寻萌 ^a ,
郭尚瑞 ^a ,
王楠 ^b ,
刘冰 ^c,d ,
王洪亮 ^a,e,,

1. 西安交通大学基础医学院病原生物学与免疫学系;
2. 西安交通大学药学院;
3. 西安交通大学第一附属医院

通讯作者： 王洪亮, hongliangwang@xjtu.edu.cn
收稿日期： 2021-11-10
录用日期： 2022-03-04

摘要

新冠病毒的流行给全球健康带来巨大影响。病毒依赖于其表面的刺突蛋白与细胞表面的受体ACE2结合而感染宿主细胞。在本研究中，我们使用了表达新冠刺突蛋白的假病毒系统分析了新冠病毒感染细胞的途径。我们发现，新冠病毒通过clathrin网格蛋白介导的内吞方式感染宿主细胞，而且磷酸肌醇通路在此过程中发挥了重要功能。此外，我们发现新冠病毒感染细胞不依赖于ACE2蛋白的胞内区及其活性催化位点。最后我们发现，传播特性更强、诱导合胞体形成能力更强的Delta突变株仍然依赖于clathrin介导的内吞感染宿主细胞。因此，本研究加深了我们对于新冠病毒感染宿主细胞的认识，同时也为针对内吞途径干预病毒感染提供了理论基础。
- 新冠病毒
- , 内吞
- , 磷酸肌醇
- , ACE2
- , 合胞体

Cytoplasmic domain and enzymatic activity of ACE2 are not required for PI4KB dependent endocytosis entry of SARS-CoV-2 into host cells

Hang Yang ^a ,
Huijun Yuan ^a ,
Xiaohui Zhao ^a ,
Meng Xun ^a ,
Shangrui Guo ^a ,
Nan Wang ^b ,
Bing Liu ^c,d ,
Hongliang Wang ^a,e,,

a Department of Pathogen Biology and Immunology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, Xi'an, 710061, China;
b School of Pharmacy, Xi'an Jiaotong University Health Science Center, Xi'an, 710061, China;
c BioBank, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, 710061, China;

Corresponding author: Hongliang Wang, hongliangwang@xjtu.edu.cn
Received Date: 10 November 2021
Accepted Date: 04 March 2022

Abstract

The recent COVID-19 pandemic poses a global health emergency. Cellular entry of the causative agent SARS-CoV-2 is mediated by its spike protein interacting with cellular receptor-human angiotensin converting enzyme 2 (ACE2). Here, by using lentivirus based pseudotypes bearing spike protein, we demonstrated that entry of SARS-CoV-2 into host cells was dependent on clathrin-mediated endocytosis, and phosphoinositides played essential roles during this process. In addition, we showed that the intracellular domain and the catalytic activity of ACE2 were not required for efficient virus entry. Finally, we showed that the current predominant Delta variant, although with high infectivity and high syncytium formation, also entered cells through clathrin-mediated endocytosis. These results provide new insights into SARS-CoV-2 cellular entry and present proof of principle that targeting viral entry could be an effective way to treat different variant infections.
- SARS-CoV-2
- , Endocytosis
- , Phosphoinositides
- , Angiotensin converting enzyme 2 (ACE2)
- , Syncytium

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