Paingankar Mandar S, Arankalle Vidya A
National Institute of Virology, Microbial Containment Complex, 130/1 Sus Road, Pashan, Pune 411 021, India.
J Biosci. 2014 Jun;39(3):389-99. doi: 10.1007/s12038-014-9436-x.
Identification and characterization of virus host interactions is an essential step for the development of novel antiviral strategies. Very few studies have been targeted towards identification of chikungunya virus (CHIKV) interacting host proteins. In current study, virus overlay protein binding assay (VOPBA) and matrix-assisted laser desorption/ ionization time of flight analysis (MALDI TOF/TOF) were employed for the identification of CHIKV binding proteins in mammalian cells. HSP70 and actin were identified as virus binding proteins in HEK-293T and Vero-E6 cells, whereas STAT-2 was identified as an additional protein in Vero-E6 cells. Pre-incubation with anti-HSP70 antibody and miRNA silencing of HSP70 significantly reduced the CHIKV production in HEK-293T and Vero-E6 cells at early time points. These results suggest that CHIKV exploits the housekeeping molecules such as actin, HSP70 and STAT-2 to establish infection in the mammalian cells.
鉴定和表征病毒与宿主的相互作用是开发新型抗病毒策略的关键步骤。针对鉴定基孔肯雅病毒(CHIKV)相互作用宿主蛋白的研究非常少。在当前研究中,采用病毒覆盖蛋白结合试验(VOPBA)和基质辅助激光解吸/电离飞行时间分析(MALDI TOF/TOF)来鉴定哺乳动物细胞中的CHIKV结合蛋白。HSP70和肌动蛋白被鉴定为HEK-293T和Vero-E6细胞中的病毒结合蛋白,而STAT-2被鉴定为Vero-E6细胞中的另一种蛋白。用抗HSP70抗体预孵育和HSP70的miRNA沉默在早期时间点显著降低了HEK-293T和Vero-E6细胞中的CHIKV产生。这些结果表明,CHIKV利用肌动蛋白、HSP70和STAT-2等管家分子在哺乳动物细胞中建立感染。