Laboratory of Protein Chemistry and Biochemistry, Department of Cell Biology, Institute of Biology, University of Brasilia, Brasilia, DF, 70910-900, Brazil.
Laboratory of Virology, Department of Cell Biology, Institute of Biology, University of Brasilia, Brasilia, DF, 70910-900, Brazil.
Parasit Vectors. 2020 Jun 10;13(1):297. doi: 10.1186/s13071-020-04167-2.
Mayaro virus (MAYV) is responsible for a mosquito-borne tropical disease with clinical symptoms similar to dengue or chikungunya virus fevers. In addition to the recent territorial expansion of MAYV, this virus may be responsible for an increasing number of outbreaks. Currently, no vaccine is available. Aedes aegypti is promiscuous in its viral transmission and thus an interesting model to understand MAYV-vector interactions. While the life-cycle of MAYV is known, the mechanisms by which this arbovirus affects mosquito host cells are not clearly understood.
After defining the best conditions for cell culture harvesting using the highest virus titer, Ae. aegypti Aag-2 cells were infected with a Brazilian MAYV isolate at a MOI of 1 in order to perform a comparative proteomic analysis of MAYV-infected Aag-2 cells by using a label-free semi-quantitative bottom-up proteomic analysis. Time-course analyses were performed at 12 and 48 h post-infection (hpi). After spectrum alignment between the triplicates of each time point and changes of the relative abundance level calculation, the identified proteins were annotated and using Gene Ontology database and protein pathways were annotated using the Kyoto Encyclopedia of Genes and Genomes.
After three reproducible biological replicates, the total proteome analysis allowed for the identification of 5330 peptides and the mapping of 459, 376 and 251 protein groups, at time 0, 12 hpi and 48 hpi, respectively. A total of 161 mosquito proteins were found to be differentially abundant during the time-course, mostly related to host cell processes, including redox metabolism, translation, energy metabolism, and host cell defense. MAYV infection also increased host protein expression implicated in viral replication.
To our knowledge, this first proteomic time-course analysis of MAYV-infected mosquito cells sheds light on the molecular basis of the viral infection process and host cell response during the first 48 hpi. Our data highlight several mosquito proteins modulated by the virus, revealing that MAYV manipulates mosquito cell metabolism for its propagation.
马雅罗病毒（MAYV）是一种蚊媒热带疾病的病原体，其临床症状与登革热或基孔肯雅热病毒发热相似。除了 MAYV 最近的领土扩张外，这种病毒可能导致越来越多的疫情爆发。目前，尚无疫苗可用。埃及伊蚊在其病毒传播中是混杂的，因此是了解 MAYV-媒介相互作用的一个有趣模型。虽然 MAYV 的生命周期是已知的，但这种虫媒病毒影响蚊子宿主细胞的机制尚不清楚。
在用最高病毒滴度确定细胞培养收获的最佳条件后，用 MOI 为 1 的巴西 MAYV 分离株感染埃及伊蚊 Aag-2 细胞，以进行用无标签半定量底向上蛋白质组学分析比较 MAYV 感染的 Aag-2 细胞。在感染后 12 和 48 小时（hpi）进行时间进程分析。在每个时间点的三个重复之间进行光谱比对并计算相对丰度水平变化后，对鉴定的蛋白质进行注释，并使用基因本体数据库注释蛋白质途径，使用京都基因与基因组百科全书注释。
经过三个可重复的生物学重复，总蛋白质组分析允许鉴定 5330 个肽和映射 459、376 和 251 个蛋白质组，分别在时间 0、12 hpi 和 48 hpi。在时间过程中，共发现 161 种蚊子蛋白丰度差异，主要与宿主细胞过程有关，包括氧化还原代谢、翻译、能量代谢和宿主细胞防御。MAYV 感染还增加了与病毒复制有关的宿主蛋白表达。
据我们所知，这是对 MAYV 感染蚊子细胞的第一个蛋白质组时间过程分析，揭示了病毒感染过程和宿主细胞在最初 48 hpi 期间的反应的分子基础。我们的数据突出了几个被病毒调节的蚊子蛋白，揭示 MAYV 操纵蚊子细胞代谢以促进其繁殖。
