Quantitative Proteomic Analysis of Chikungunya Virus-Infected Aedes aegypti Reveals Proteome Modulations Indicative of Persistent Infection

The mosquito-borne chikungunya virus (CHIKV) poses a threat to human health in tropical countries throughout the world. The molecular interactions of CHIKV with its mosquito vector Aedes aegypti are not fully understood. Following oral acquisition of CHIKV via salinemeals, we analyzed changes in the proteome of Ae. aegypti in 12 h intervals by label-free quantification using a timsTOF Pro mass spectrometer. For each of the seven time points, between 2647 and 3167 proteins were identified among CHIKV-infected and noninfected mosquito samples, and fewer than 6% of those identified proteins were affected by the virus. Functional enrichment analysis revealed that the three pathways, Endocytosis, Oxidative phosphorylation, and Ribosome biogenesis, were enriched during CHIKV infection. On the other hand, three pathways of the cellular RNA machinery and five metabolism related pathways were significantly attenuated in the CHIKV-infected samples. Furthermore, proteins associated with cytoskeleton and vesicular transport, as well as various serine-type endopeptidases and metallo-proteinases, were modulated in the presence of CHIKV. Our study reveals biological pathways and novel proteins interacting with CHIKV in the mosquito. Overall, CHIKV infection caused minor changes to the mosquito proteome demonstrating a high level of adaption between the vector and the virus, essentially coexisting in a nonpathogenic relationship. The mass spectrometry data have been deposited to the MassIVE repository (https://massive.ucsd.edu/ProteoSAFe/dataset.jsp?task=abfd14f7015243c69854731998d55df1) with the data set identifier MSV000085115.经蚊虫传播的基孔肯雅病毒（chikungunya virus, CHIKV）对全球热带地区的人类健康构成威胁。目前学界尚未完全阐明CHIKV与其蚊虫媒介埃及伊蚊（Aedes aegypti）之间的分子互作机制。通过盐溶液饲喂经口获取CHIKV后，本研究采用基于timsTOF Pro质谱仪的无标记定量技术，以12小时间隔对埃及伊蚊的蛋白质组变化进行分析。在7个时间点的每一个中，CHIKV感染组与非感染组蚊虫样本中共鉴定出2647至3167种蛋白质，其中仅有不到6%的鉴定蛋白受病毒调控。功能富集分析显示，内吞作用（Endocytosis）、氧化磷酸化（Oxidative phosphorylation）以及核糖体生物发生（Ribosome biogenesis）这三条通路在CHIKV感染期间显著富集。另一方面，CHIKV感染样本中，细胞RNA机器相关的三条通路以及五条代谢相关通路均显著下调。此外，在CHIKV感染条件下，与细胞骨架、囊泡运输相关的蛋白质，以及多种丝氨酸型内肽酶和金属蛋白酶均发生了表达调控。本研究揭示了蚊虫体内与CHIKV互作的生物学通路及新型蛋白质。总体而言，CHIKV感染仅对蚊虫蛋白质组造成轻微改变，这表明媒介与病毒之间存在高度的适应性，二者本质上以非致病性关系共存。本研究的质谱数据已上传至MassIVE数据库（https://massive.ucsd.edu/ProteoSAFe/dataset.jsp?task=abfd14f7015243c69854731998d55df1），数据集标识符为MSV000085115。