Mishra Nimisha, Chaudhary Yash, Chaudhary Sakshi, Singh Anjali, Srivastava Priyanshu, Sunil Sujatha
Vector Borne Disease Group, International Centre for Genetic Engineering and Biotechnology, New Delhi 110067, India.
TERI School of Advanced Studies (TERI-SAS), New Delhi 110070, India.
Int J Mol Sci. 2025 Jul 16;26(14):6832. doi: 10.3390/ijms26146832.
Chikungunya virus (CHIKV), a mosquito-borne alphavirus, has re-emerged, causing widespread outbreaks and a significant clinical burden. Despite advances in virology, the molecular mechanisms governing CHIKV's interaction with host cells remain poorly understood. In this study, we aimed to identify novel host protein interactors of the CHIKV nonstructural protein 3 (nsP3), a critical component of the viral replication complex, using mass spectrometry-based proteomic profiling in liver-derived Huh7 cells. Co-immunoprecipitation followed by LC-MS/MS identified a wide array of host proteins associated with nsP3, revealing 52 proteins classified as high-confidence (FDR of 1%, and unique peptides > 2) CHIKV-specific interactors. A bioinformatic analysis using STRING and Cytoscape uncovered interaction networks enriched in metabolic processes, RNA processing, translation regulation, cellular detoxification, stress responses, and immune signaling pathways. A subcellular localization analysis showed that many interactors reside in the cytosol, while others localize to the nucleus, nucleolus, and mitochondria. Selected novel host protein interactions were validated through co-immunoprecipitation and immunofluorescence assays. Our findings provide new insights into the host cellular pathways hijacked by CHIKV and highlight potential targets for therapeutic intervention. This is the first report mapping direct nsP3-host protein interactions in Huh7 cells during CHIKV infection.
基孔肯雅病毒（CHIKV）是一种由蚊子传播的甲病毒，现已再次出现，导致广泛的疫情爆发和严重的临床负担。尽管病毒学取得了进展，但对于CHIKV与宿主细胞相互作用的分子机制仍知之甚少。在本研究中，我们旨在利用基于质谱的蛋白质组学分析方法，在源自肝脏的Huh7细胞中鉴定CHIKV非结构蛋白3（nsP3）的新型宿主蛋白相互作用分子，nsP3是病毒复制复合体的关键组成部分。通过免疫共沉淀结合液相色谱-串联质谱（LC-MS/MS）鉴定出了大量与nsP3相关的宿主蛋白，揭示了52种被归类为高可信度（错误发现率为1%，且独特肽段>2）的CHIKV特异性相互作用分子。使用STRING和Cytoscape进行的生物信息学分析揭示了富含代谢过程、RNA加工、翻译调控、细胞解毒、应激反应和免疫信号通路的相互作用网络。亚细胞定位分析表明，许多相互作用分子位于细胞质中，而其他分子则定位于细胞核、核仁及线粒体。通过免疫共沉淀和免疫荧光试验验证了所选的新型宿主蛋白相互作用。我们的研究结果为CHIKV劫持的宿主细胞途径提供了新的见解，并突出了治疗干预的潜在靶点。这是首篇关于CHIKV感染期间Huh7细胞中nsP3与宿主蛋白直接相互作用图谱的报告。
