De Caluwé Lien, Coppens Sandra, Vereecken Katleen, Daled Simon, Dhaenens Maarten, Van Ostade Xaveer, Deforce Dieter, Ariën Kevin K, Bartholomeeusen Koen
Virology Unit, Biomedical Sciences, Institute of Tropical Medicine Antwerp, Antwerp, Belgium.
Laboratory for Pharmaceutical Biotechnology, University of Ghent, Ghent, Belgium.
Front Microbiol. 2021 Feb 25;12:615165. doi: 10.3389/fmicb.2021.615165. eCollection 2021.
Chikungunya virus (CHIKV) is an arbovirus with a global spread and significant public health impact. It is a positive stranded RNA alphavirus belonging to the family. However, many questions about the replication cycle of CHIKV remain unanswered. The entry process of CHIKV is not completely understood nor are the associated virus-receptor interactions fully identified. Here, we designed an affinity purification mass spectrometry coupled approach that allowed the identification of factors that facilitate entry of CHIKV in human cells. The identified entry factors were further validated using CRISPR/Cas9. In HEK293T cells we identified the CD147 protein complex as an entry factor for CHIKV. We further showed the involvement of the CD147 protein complex in the replication cycle of related alphaviruses. Interestingly, CD147 contains similar protein domains as the previously identified alphavirus entry factor MXRA8.
基孔肯雅病毒(CHIKV)是一种虫媒病毒,在全球传播,对公共卫生有重大影响。它是一种正链RNA甲病毒,属于该家族。然而,关于CHIKV复制周期的许多问题仍未得到解答。CHIKV的进入过程尚未完全了解,相关的病毒-受体相互作用也未完全确定。在这里,我们设计了一种亲和纯化质谱联用方法,该方法能够鉴定促进CHIKV进入人细胞的因子。使用CRISPR/Cas9对鉴定出的进入因子进行了进一步验证。在HEK293T细胞中,我们鉴定出CD147蛋白复合物是CHIKV的进入因子。我们进一步证明了CD147蛋白复合物参与了相关甲病毒的复制周期。有趣的是,CD147包含与先前鉴定的甲病毒进入因子MXRA8相似的蛋白结构域。