Fongsaran Chanida, Jirakanwisal Krit, Kuadkitkan Atichat, Wikan Nitwara, Wintachai Phitchayapak, Thepparit Chutima, Ubol Sukathida, Phaonakrop Narumon, Roytrakul Sittiruk, Smith Duncan R
Institute of Molecular Biosciences, Mahidol University, Salaya Campus, 25/25 Phuttamonthol Sai 4, Nakhon Pathom, Bangkok, 73170, Thailand.
Arch Virol. 2014 Dec;159(12):3353-64. doi: 10.1007/s00705-014-2210-4. Epub 2014 Aug 29.
Chikungunya virus (CHIKV), the virus responsible for the disease chikungunya fever in humans, is transmitted by Aedes mosquitoes. While significant progress has been made in understanding the process by which CHIKV enters into mammalian cells, far less progress has been made in understanding the CHIKV entry process in insect cells. This study sought to identify mosquito-cell-expressed CHIKV-binding proteins through a combination of virus overlay protein binding assays (VOPBA) and mass spectroscopy. A 50-kDa CHIKV-binding protein was identified as the ATP synthase β subunit (ATPSβ). Co-immunoprecipitation studies confirmed the interaction, and colocalization analysis showed cell-surface and intracellular co-localization between CHIKV and ATPSβ. Both antibody inhibition and siRNA-mediated downregulation experiments targeted to ATPSβ showed a significant reduction in viral entry and virus production. These results suggest that ATPSβ is a CHIKV-binding protein capable of mediating the entry of CHIKV into insect cells.
基孔肯雅病毒（CHIKV）是导致人类基孔肯雅热的病毒，通过伊蚊传播。虽然在了解CHIKV进入哺乳动物细胞的过程方面取得了重大进展，但在了解CHIKV进入昆虫细胞的过程方面进展甚微。本研究旨在通过病毒覆盖蛋白结合试验（VOPBA）和质谱相结合的方法，鉴定蚊子细胞表达的CHIKV结合蛋白。一种50 kDa的CHIKV结合蛋白被鉴定为ATP合酶β亚基（ATPSβ）。免疫共沉淀研究证实了这种相互作用，共定位分析显示CHIKV与ATPSβ在细胞表面和细胞内共定位。针对ATPSβ的抗体抑制和siRNA介导的下调实验均显示病毒进入和病毒产生显著减少。这些结果表明，ATPSβ是一种能够介导CHIKV进入昆虫细胞的CHIKV结合蛋白。
