Systematically Quantitative Analysis Of Zika Virus And Host Interactions

Viruses are a type of stringent intracellular parasite,and thus the interaction between the virus and the host is the focus of virological study.Current research methods include high-throughput screening techniques,such as whole-genome knockdown or knockout and ectopic overexpression,or systematic quantitative studies using omics methods.In recent years,Zika virus has received worldwide attention because of its nervous system-related diseases such as microcephaly.There is no approved drug or vaccine for Zika virus.Therefore,understanding the interaction between Zika virus and the host facilitates the understanding of the virus itself and the design of drug targets.Since there is currently no relevant research on the quantitative proteomics of Zika virus and host,we performed a systematic quantitative analysis of Zika virus-infected C6/36 and human glial cells U251 using iTRAQ-based quantitative proteomics.In quantitative proteomics of ZIKV-infected mosquito cells C6/36,we identified 3,544 host proteins,of which 200 were differentially expressed.Further bioinformatics analysis found that the biological processes of differentially expressed proteins mainly cluster into material energy metabolism processes,mitochondrial related components,and transcription-translation-related processes.In functional assays,it was found that ZIKV can upregulate KXJ75878(and CHCHD2 homologue)proteins that inhibit interferon function to promote viral replication after infection with C6/36 cells.Finally,we found that the ubiquitin-proteasome inhibitor Bortezomib can effectively inhibit viral replication at the nM level in cell experiments,and it can also effectively inhibit Zika virus load and alleviate pathological injury in animal experiments.In the quantitative proteomics results of ZIKV-infected human glial cell U251,we identified a total of 5,994 host proteins.The number of differentially expressed proteins at four different time points was 63,41,90,and 487,respectively.Bioinformatics analysis showed that differentially expressed proteins mainly clustered in mitochondria-associated proteins,endoplasmic reticulum-associated proteins,antiviral innate immune-related proteins,and intracellular homeostasis-related proteins.We therefore performed live-cell imaging of the endoplasmic reticulum and lysosome structure and found that the structure of the endoplasmic reticulum is rearranged during the mid-stage of viral infection,and the endoplasmic reticulum is degraded by lysosomes at the end of the period,while the lysosomal inhibitor BFA can effectively relieve endoplasmic reticulum degradation process.Finally,we also verified that lysosomal inhibitors BFA as well as CQ can significantly inhibit Zika virus during virus entry and mid-release process.In summary,we systematically studied the interaction between Zika virus and two different hosts using iTRAQ-based quantitative proteomics method.A systematic comparison of the similarities and differences between the same virus in different hosts can help us fully understand Zika.The different replication of the virus between the proliferating host and the final host has important guiding significance for a comprehensive understanding of Zika virus-related biological information,drug target design and subsequent functional analysis experiments.