Carcinoembryonic Antigen-Related Cell Adhesion Molecule 5 Is an Important Surface Attachment Factor That Facilitates Entry of Middle East Respiratory Syndrome Coronavirus
Carcinoembryonic Antigen-Related Cell Adhesion Molecule 5 Is an Important Surface Attachment Factor That Facilitates Entry of Middle East Respiratory Syndrome Coronavirus
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癌胚抗原相关细胞黏附分子5是促进中东呼吸综合征冠状病毒入侵的重要表面黏附因子
DOI：
10.1128/jvi.01133-16
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发表时间：
2016-10-01
影响因子：
5.4
通讯作者：
Yuen, Kwok-Yung
中科院分区：
文献类型：
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作者：
Chan, Che-Man;Chu, Hin;Yuen, Kwok-Yung
冠状病毒的刺突蛋白能够与广泛的细胞靶点结合，这有助于冠状病毒的广泛种属嗜性。先前的报道已经证明中东呼吸综合征冠状病毒（MERSCoV）主要利用二肽基肽酶4（DPP 4）进入细胞。然而，MERS-CoV刺突蛋白的可能增加MERS-CoV感染的其他细胞结合靶点尚未进一步探索。在目前的研究中，使用病毒覆盖蛋白结合试验（VOPBA），我们确定癌胚抗原相关的细胞粘附分子5（CEACAM 5）作为MERS-CoV的新的细胞表面结合靶点。CEACAM 5与MERS-CoV的刺突蛋白在过表达和内源性环境中共免疫沉淀。用抗CEACAM 5抗体、重组CEACAM 5蛋白或CEACAM 5的小干扰RNA（siRNA）敲低破坏CEACAM 5与MERS-CoV刺突之间的相互作用显著抑制MERS-CoV的进入。CEACAM 5的重组表达不会使非允许性幼仓鼠肾（BHK 21）细胞对MERS-CoV感染敏感。相反，CEACAM 5过表达显著增强了MERS-CoV与BHK 21细胞的附着。更重要的是，当CEACAM 5在允许细胞中过表达时，MERS-CoV的进入增加，这表明CEACAM 5可以与DPP 4一起促进MERS-CoV进入，尽管不能独立地支持MERS-CoV进入。综上所述，我们的研究结果确定CEACAM 5作为MERS-CoV的一种新的细胞表面结合靶点，通过增加病毒与宿主细胞表面的附着来促进MERS-CoV感染。重要中东呼吸综合征冠状病毒（MERS-CoV）感染与所有已知的人类致病性冠状病毒中最高的死亡率相关。目前，没有针对MERSCoV感染的批准的疫苗或治疗剂。癌胚抗原相关细胞粘附分子5（CEACAM 5）作为MERS-CoV细胞表面结合新靶点的发现，为MERS-CoV的细胞结合生物学研究提供了新的思路。重要的是，CEACAM 5可以通过充当附着因子来增强MERS-CoV的进入。在这方面，CEACAM 5可以作为一个新的目标，除了二肽基肽酶-4（DPP 4），在开发MERS-CoV的抗病毒策略。
The spike proteins of coronaviruses are capable of binding to a wide range of cellular targets, which contributes to the broad species tropism of coronaviruses. Previous reports have demonstrated that Middle East respiratory syndrome coronavirus (MERSCoV) predominantly utilizes dipeptidyl peptidase 4 (DPP4) for cell entry. However, additional cellular binding targets of the MERS-CoV spike protein that may augment MERS-CoV infection have not been further explored. In the current study, using the virus overlay protein binding assay (VOPBA), we identified carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5) as a novel cell surface binding target of MERS-CoV. CEACAM5 coimmunoprecipitated with the spike protein of MERS-CoV in both overexpressed and endogenous settings. Disrupting the interaction between CEACAM5 and MERS-CoV spike with anti-CEACAM5 antibody, recombinant CEACAM5 protein, or small interfering RNA (siRNA) knockdown of CEACAM5 significantly inhibited the entry of MERS-CoV. Recombinant expression of CEACAM5 did not render nonpermissive baby hamster kidney (BHK21) cells susceptible to MERS-CoV infection. Instead, CEACAM5 overexpression significantly enhanced the attachment of MERS-CoV to the BHK21 cells. More importantly, the entry of MERS-CoV was increased when CEACAM5 was overexpressed in permissive cells, which suggested that CEACAM5 could facilitate MERS-CoV entry in conjunction with DPP4 despite not being able to support MERS-CoV entry independently. Taken together, the results of our study identified CEACAM5 as a novel cell surface binding target of MERS-CoV that facilitates MERS-CoV infection by augmenting the attachment of the virus to the host cell surface.IMPORTANCEInfection with the Middle East respiratory syndrome coronavirus (MERS-CoV) is associated with the highest mortality rate among all known human-pathogenic coronaviruses. Currently, there are no approved vaccines or therapeutics against MERSCoV infection. The identification of carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5) as a novel cell surface binding target of MERS-CoV advanced our knowledge on the cell binding biology of MERS-CoV. Importantly, CEACAM5 could potentiate the entry of MERS-CoV by functioning as an attachment factor. In this regard, CEACAM5 could serve as a novel target, in addition to dipeptidyl peptidase-4 (DPP4), in the development of antiviral strategies for MERS-CoV.