Department of Molecular and Cell Biology, Centro Nacional de Biotecnología (CNB-CSIC), Madrid, Spain.
Department of Infectious Diseases and Global Health, Animal Health Research Center (CISA), National Institute of Research, Agricultural and Food Technology (INIA-CSIC), Valdeolmos, Madrid, Spain.
mBio. 2023 Feb 28;14(1):e0313622. doi: 10.1128/mbio.03136-22. Epub 2023 Jan 10.
Coronaviruses (CoVs) of genera α, β, γ, and δ encode proteins that have a PDZ-binding motif (PBM) consisting of the last four residues of the envelope (E) protein (PBM core). PBMs may bind over 400 cellular proteins containing PDZ domains (an acronym formed by the combination of the first letter of the names of the three first proteins where this domain was identified), making them relevant for the control of cell function. Three highly pathogenic human CoVs have been identified to date: severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV), and SARS-CoV-2. The PBMs of the three CoVs were virulence factors. SARS-CoV mutants in which the E protein PBM core was replaced by the E protein PBM core from virulent or attenuated CoVs were constructed. These mutants showed a gradient of virulence, depending on whether the alternative PBM core introduced was derived from a virulent or an attenuated CoV. Gene expression patterns in the lungs of mice infected with SARS-CoVs encoding each of the different PBMs were analyzed by RNA sequencing of infected lung tissues. E protein PBM of SARS-CoV and SARS-CoV-2 dysregulated gene expression related to ion transport and cell homeostasis. Decreased expression of cystic fibrosis transmembrane conductance regulator (CFTR) mRNA, essential for alveolar edema resolution, was shown. Reduced CFTR mRNA levels were associated with edema accumulation in the alveoli of mice infected with SARS-CoV and SARS-CoV-2. Compounds that increased CFTR expression and activity, significantly reduced SARS-CoV-2 growth in cultured cells and protected against mouse infection, suggesting that E protein virulence is mediated by a decreased CFTR expression. Three highly pathogenic human CoVs have been identified: SARS-CoV, MERS-CoV, and SARS-CoV-2. The E protein PBMs of these three CoVs were virulence factors. Gene expression patterns associated with the different PBM motifs in the lungs of infected mice were analyzed by deep sequencing. E protein PBM motif of SARS-CoV and SARS-CoV-2 dysregulated the expression of genes related to ion transport and cell homeostasis. A decrease in the mRNA expression of the cystic fibrosis transmembrane conductance regulator (CFTR), which is essential for edema resolution, was observed. The reduction of CFTR mRNA levels was associated with edema accumulation in the lungs of mice infected with SARS-CoV-2. Compounds that increased the expression and activity of CFTR drastically reduced the production of SARS-CoV-2 and protected against its infection in a mice model. These results allowed the identification of cellular targets for the selection of antivirals.
严重急性呼吸系统综合征冠状病毒（SARS-CoV）和中东呼吸系统综合征冠状病毒（MERS-CoV），以及 SARS-CoV-2。这三种冠状病毒的 PBM 是毒力因子。构建了 E 蛋白 PBM 核心由毒力或减毒冠状病毒的 E 蛋白 PBM 核心取代的 SARS-CoV 突变体。这些突变体显示出梯度的毒力，这取决于引入的替代 PBM 核心是来自毒力还是减毒的冠状病毒。通过感染肺组织的 RNA 测序分析了感染 SARS-CoV 的小鼠肺部的基因表达模式，这些 SARS-CoV 编码不同的 PBM。SARS-CoV 和 SARS-CoV-2 的 E 蛋白 PBM 扰乱了与离子转运和细胞内稳态相关的基因表达。囊性纤维化跨膜电导调节因子（CFTR）mRNA 的表达减少，这对肺泡水肿的消退至关重要。CFTR mRNA 水平的降低与感染 SARS-CoV 和 SARS-CoV-2 的小鼠肺泡中的水肿积累有关。增加 CFTR 表达和活性的化合物显著减少了 SARS-CoV-2 在培养细胞中的生长，并防止了小鼠感染，这表明 E 蛋白的毒力是通过降低 CFTR 的表达来介导的。
SARS-CoV、MERS-CoV 和 SARS-CoV-2。这些三种冠状病毒的 E 蛋白 PBM 是毒力因子。通过深度测序分析了感染小鼠肺部不同 PBM 模式相关的基因表达模式。SARS-CoV 和 SARS-CoV-2 的 E 蛋白 PBM 扰乱了与离子转运和细胞内稳态相关的基因表达。观察到囊性纤维化跨膜电导调节因子（CFTR）mRNA 表达减少，这对水肿消退至关重要。CFTR mRNA 水平的降低与感染 SARS-CoV-2 的小鼠肺部的水肿积累有关。增加 CFTR 表达和活性的化合物可显著减少 SARS-CoV-2 的产生，并在小鼠模型中防止其感染。这些结果为抗病毒药物的选择确定了细胞靶标。
