Zhang Qiangmin, Gefter Julia, Sneddon W Bruce, Mamonova Tatyana, Friedman Peter A
Laboratory for GPCR Biology, Department of Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh PA15261, USA.
iScience. 2021 Jul 23;24(7):102770. doi: 10.1016/j.isci.2021.102770. Epub 2021 Jun 24.
SARS-CoV-2 is responsible for the global COVID-19 pandemic. Angiotensin converting enzyme 2 (ACE2) is the membrane-delimited receptor for SARS-CoV-2. Lung, intestine, and kidney, major sites of viral infection, express ACE2 that harbors an intracellular, carboxy-terminal PDZ-recognition motif. These organs prominently express the PDZ protein Na/H exchanger regulatory factor-1 (NHERF1). Here, we report NHERF1 tethers ACE2 and augments SARS-CoV-2 cell entry. ACE2 directly binds both NHERF1 PDZ domains. Disruption of either NHERF1 PDZ core-binding motif or the ACE2 PDZ recognition sequence eliminates interaction. Proximity ligation assays establish that ACE2 and NHERF1 interact at constitutive expression levels in human lung and intestine cells. Ablating ACE2 interaction with NHERF1 accelerated SARS-CoV-2 cell entry. Conversely, elimination of the ACE2 C-terminal PDZ-binding motif decreased ACE2 membrane residence and reduced pseudotyped virus entry. We conclude that the PDZ interaction of ACE2 with NHERF1 facilitates SARS-CoV-2 internalization. β-Arrestin is likely indispensable, as with G protein-coupled receptors.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）引发了全球新型冠状病毒肺炎（COVID-19）大流行。血管紧张素转换酶2（ACE2）是SARS-CoV-2的膜结合受体。肺、肠道和肾脏是病毒感染的主要部位，表达含有细胞内羧基末端PDZ识别基序的ACE2。这些器官显著表达PDZ蛋白钠/氢交换调节因子1（NHERF1）。在此，我们报告NHERF1与ACE2相连并增强SARS-CoV-2进入细胞。ACE2直接结合NHERF1的两个PDZ结构域。破坏NHERF1的PDZ核心结合基序或ACE2的PDZ识别序列会消除相互作用。邻近连接分析表明，ACE2和NHERF1在人肺和肠道细胞的组成型表达水平上相互作用。消除ACE2与NHERF1的相互作用会加速SARS-CoV-2进入细胞。相反，去除ACE2的C末端PDZ结合基序会降低ACE2在膜上的停留时间并减少假型病毒的进入。我们得出结论，ACE2与NHERF1的PDZ相互作用促进了SARS-CoV-2的内化。与G蛋白偶联受体一样，β-抑制蛋白可能不可或缺。
