Comprehensive Quantitative Proteome Analysis of Aedes aegypti Identifies Proteins and Pathways Involved in Wolbachia pipientis and Zika Virus Interference Phenomenon

Zika virus (ZIKV) is a global public health emergency due to its association with microcephaly, Guillain-Barré syndrome, neuropathy, and myelitis in children and adults. A total of 87 countries have had evidence of autochthonous mosquito-borne transmission of ZIKV, distributed across four continents, and no antivirus therapy or vaccines are available. Therefore, several strategies have been developed to target the main mosquito vector, Aedes aegypti, to reduce the burden of different arboviruses. Among such strategies, the use of the maternally-inherited endosymbiont Wolbachia pipientis has been applied successfully to reduce virus susceptibility and decrease transmission. However, the mechanisms by which Wolbachia orchestrate resistance to ZIKV infection remain to be elucidated. In this study, we apply isobaric labeling quantitative mass spectrometry (MS)-based proteomics to quantify proteins and identify pathways altered during ZIKV infection; Wolbachia infection; co-infection with Wolbachia/ZIKV in the A. aegypti heads and salivary glands. We show that Wolbachia regulates proteins involved in reactive oxygen species production, regulates humoral immune response, and antioxidant production. The reduction of ZIKV polyprotein in the presence of Wolbachia in mosquitoes was determined by MS and corroborates the idea that Wolbachia helps to block ZIKV infections in A. aegypti. The present study offers a rich resource of data that may help to elucidate mechanisms by which Wolbachia orchestrate resistance to ZIKV infection in A. aegypti, and represents a step further on the development of new targeted methods to detect and quantify ZIKV and Wolbachia directly in complex tissues.

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Comprehensive quantitative proteome analysis of Aedes aegypti identifies proteins and pathways involved in Wolbachia pipientis and Zika virus interference phenomenon

10.1101/2020.12.12.422506 ◽

2020 ◽

Author(s):

Michele Martins ◽

Luis Felipe Costa Ramos ◽

Jimmy Rodriguez Murillo ◽

André Torres ◽

Stephanie Serafim de Carvalho ◽

...

Keyword(s):

Mass Spectrometry ◽

Immune Response ◽

Aedes Aegypti ◽

Humoral Immune Response ◽

Zika Virus ◽

Mosquito Vector ◽

Ros Production ◽

Zikv Infection ◽

Wolbachia Pipientis ◽

Humoral Immune

ABSTRACTZika virus is a global public health emergency due to its association with microcephaly, Guillain-Barré syndrome, neuropathy, and myelitis in children and adults. A total of 87 countries have had evidence of autochthonous mosquito-borne transmission of Zika virus, distributed across four continents, and no antivirus therapy or vaccines are available. Therefore, several strategies have been developed to target the main mosquito vector, Aedes aegypti, to reduce the burden of different arboviruses. Among such strategies, the use of the maternally-inherited endosymbiont Wolbachia pipientis has been applied successfully to reduce virus susceptibility and decrease transmission. However, the mechanisms by which Wolbachia orchestrate resistance to ZIKV infection remain to be elucidated. In this study, we apply isobaric labeling quantitative mass spectrometry-based proteomics to quantify proteins and identify pathways altered during ZIKV infection; Wolbachia infection; co-infection with Wolbachia/ZIKV in the Ae. aegypti heads and salivary glands. We show that Wolbachia regulates proteins involved in ROS production, regulates humoral immune response, and antioxidant production. The reduction of ZIKV polyprotein in the presence of Wolbachia in mosquitoes was determined by mass spectrometry and corroborates the idea that Wolbachia helps to block ZIKV infections in Ae. aegypti. The present study offers a rich resource of data that may help to elucidate mechanisms by which Wolbachia orchestrate resistance to ZIKV infection in Ae. aegypti, and represents a step further on the development of new targeted methods to detect and quantify ZIKV and Wolbachia directly in complex tissues.HighlightsThe abundance of ZIKV polyprotein is reduced in the presence of WolbachiaShotgun proteomics quantifies ZIKV and Wolbachia proteins directly in tissuesWolbachia regulates proteins involved in ROS productionWolbachia regulates humoral immune response and antioxidant productionMetabolism and detoxification processes were associated with mono infections

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Global transcriptome analysis of Aedes aegypti mosquitoes in response to Zika virus infection

10.1101/179416 ◽

2017 ◽

Cited By ~ 1

Author(s):

Kayvan Etebari ◽

Shivanand Hegde ◽

Miguel A Saldaña ◽

Steven G Widen ◽

Thomas G Wood ◽

...

Keyword(s):

Virus Infection ◽

Aedes Aegypti ◽

Dengue Virus ◽

Zika Virus ◽

Target Genes ◽

World Health ◽

Mosquito Vector ◽

Zikv Infection ◽

Zika Virus Infection ◽

Non Coding Rnas

AbstractZika virus (ZIKV) of the Flaviviridae family is a recently emerged mosquito-borne virus that has been implicated in the surge of the number of microcephaly instances in South America. The recent rapid spread of the virus led to its declaration as a global health emergency by the World Health Organization. The virus is transmitted mainly by the mosquito Aedes aegypti that also vectors dengue virus, however little is known about the interactions of the virus with the mosquito vector. In this study, we investigated the transcriptome profiles of whole Ae. aegypti mosquitoes in response to ZIKV infection at 2, 7, and 14 days post-infection using RNA-Seq. Results showed changes in the abundance of a large number of transcripts at each time point following infection, with 18 transcripts commonly changed among the three time points. Gene ontology analysis revealed that most of the altered genes are involved in metabolic process, cellular process and proteolysis. In addition, 486 long intergenic non-coding RNAs were identified that were altered upon ZIKV infection. Further, we found correlational changes of a number of potential mRNA target genes with that of altered host microRNAs. The outcomes provide a basic understanding of Ae. aegypti responses to ZIKV and helps to determine host factors involved in replication or mosquito host anti-viral response against the virus.ImportanceVector-borne viruses pose great risks on human health. Zika virus has recently emerged as a global threat, rapidly expanding its distribution. Understanding the interactions of the virus with mosquito vectors at the molecular level is vital for devising new approaches in inhibiting virus transmission. In this study, we embarked on analyzing the transcriptional response of Aedes aegypti mosquitoes to Zika virus infection. Results showed large changes both in coding and long non-coding RNAs. Analysis of these genes showed similarities with other flaviviruses, including dengue virus, which is transmitted by the same mosquito vector. The outcomes provide a global picture of changes in the mosquito vector in response to Zika virus infection.

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Molecular Responses to the Zika Virus in Mosquitoes

Pathogens ◽

10.3390/pathogens7020049 ◽

2018 ◽

Vol 7 (2) ◽

pp. 49 ◽

Cited By ~ 4

Author(s):

Catalina Alfonso-Parra ◽

Frank Avila

Keyword(s):

Aedes Aegypti ◽

Aedes Albopictus ◽

Birth Defects ◽

Molecular Interactions ◽

Zika Virus ◽

Neurological Complications ◽

Mosquito Vector ◽

Zikv Infection ◽

Molecular Responses ◽

The Pacific

The Zika virus (ZIKV), originally discovered in 1947, did not become a major concern until the virus swept across the Pacific and into the Americas in the last decade, bringing with it news of neurological complications and birth defects in ZIKV affected areas. This prompted researchers to dissect the molecular interactions between ZIKV and the mosquito vector in an attempt to better understand not only the changes that occur upon infection, but to also identify molecules that may potentially enhance or suppress a mosquito’s ability to become infected and/or transmit the virus. Here, we review what is currently known regarding ZIKV-mosquito molecular interactions, focusing on ZIKV infection of Aedes aegypti and Aedes albopictus, the primary species implicated in transmitting ZIKV during the recent outbreaks.

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Analysis of Zika virus capsid-Aedes aegypti mosquito interactome reveals pro-viral host factors critical for establishing infection

Nature Communications ◽

10.1038/s41467-021-22966-8 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Rommel J. Gestuveo ◽

Jamie Royle ◽

Claire L. Donald ◽

Douglas J. Lamont ◽

Edward C. Hutchinson ◽

...

Keyword(s):

Aedes Aegypti ◽

Protein Interactions ◽

Zika Virus ◽

Label Free ◽

Protein Protein Interactions ◽

Zikv Infection ◽

Ubiquitin Protein Ligase ◽

Mosquito Cells ◽

Endoplasmic Reticulum Protein ◽

Arboviral Diseases

AbstractThe escalating global prevalence of arboviral diseases emphasizes the need to improve our understanding of their biology. Research in this area has been hindered by the lack of molecular tools for studying virus-mosquito interactions. Here, we develop an Aedes aegypti cell line which stably expresses Zika virus (ZIKV) capsid proteins in order to study virus-vector protein-protein interactions through quantitative label-free proteomics. We identify 157 interactors and show that eight have potentially pro-viral activity during ZIKV infection in mosquito cells. Notably, silencing of transitional endoplasmic reticulum protein TER94 prevents ZIKV capsid degradation and significantly reduces viral replication. Similar results are observed if the TER94 ortholog (VCP) functioning is blocked with inhibitors in human cells. In addition, we show that an E3 ubiquitin-protein ligase, UBR5, mediates the interaction between TER94 and ZIKV capsid. Our study demonstrates a pro-viral function for TER94/VCP during ZIKV infection that is conserved between human and mosquito cells.

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Sequential Infection of Aedes aegypti Mosquitoes with Chikungunya Virus and Zika Virus Enhances Early Zika Virus Transmission

Insects ◽

10.3390/insects9040177 ◽

2018 ◽

Vol 9 (4) ◽

pp. 177 ◽

Cited By ~ 12

Author(s):

Tereza Magalhaes ◽

Alexis Robison ◽

Michael Young ◽

William Black ◽

Brian Foy ◽

...

Keyword(s):

Aedes Aegypti ◽

Blood Meal ◽

Zika Virus ◽

Chikungunya Virus ◽

Vector Competence ◽

Virus Transmission ◽

Mosquito Vector ◽

Molecular Aspects ◽

Virus Interactions ◽

Sequential Infection

In urban settings, chikungunya, Zika, and dengue viruses are transmitted by Aedes aegypti mosquitoes. Since these viruses co-circulate in several regions, coinfection in humans and vectors may occur, and human coinfections have been frequently reported. Yet, little is known about the molecular aspects of virus interactions within hosts and how they contribute to arbovirus transmission dynamics. We have previously shown that Aedes aegypti exposed to chikungunya and Zika viruses in the same blood meal can become coinfected and transmit both viruses simultaneously. However, mosquitoes may also become coinfected by multiple, sequential feeds on single infected hosts. Therefore, we tested whether sequential infection with chikungunya and Zika viruses impacts mosquito vector competence. We exposed Ae. aegypti mosquitoes first to one virus and 7 days later to the other virus and compared infection, dissemination, and transmission rates between sequentially and single infected groups. We found that coinfection rates were high after sequential exposure and that mosquitoes were able to co-transmit both viruses. Surprisingly, chikungunya virus coinfection enhanced Zika virus transmission 7 days after the second blood meal. Our data demonstrate heterologous arbovirus synergism within mosquitoes, by unknown mechanisms, leading to enhancement of transmission under certain conditions.

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Vector Competence: What Has Zika Virus Taught Us?

Viruses ◽

10.3390/v11090867 ◽

2019 ◽

Vol 11 (9) ◽

pp. 867 ◽

Cited By ~ 14

Author(s):

Sasha R. Azar ◽

Scott C. Weaver

Keyword(s):

Experimental Design ◽

Zika Virus ◽

Vector Competence ◽

Large Body ◽

Large Data ◽

Mosquito Vector ◽

Zikv Infection ◽

Data Set ◽

Short Period ◽

Meta Analyses

The unprecedented outbreak of Zika virus (ZIKV) infection in the Americas from 2015 to 2017 prompted the publication of a large body of vector competence data in a relatively short period of time. Although differences in vector competence as a result of disparities in mosquito populations and viral strains are to be expected, the limited competence of many populations of the urban mosquito vector, Aedes aegypti, from the Americas (when its susceptibility is viewed relative to other circulating/reemerging mosquito-borne viruses such as dengue (DENV), yellow fever (YFV), and chikungunya viruses (CHIKV)) has proven a paradox for the field. This has been further complicated by the lack of standardization in the methodologies utilized in laboratory vector competence experiments, precluding meta-analyses of this large data set. As the calls for the standardization of such studies continue to grow in number, it is critical to examine the elements of vector competence experimental design. Herein, we review the various techniques and considerations intrinsic to vector competence studies, with respect to contemporary findings for ZIKV, as well as historical findings for other arboviruses, and discuss potential avenues of standardization going forward.

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Low Aedes aegypti Vector Competence for Zika Virus from Viremic Rhesus Macaques

Viruses ◽

10.3390/v12121345 ◽

2020 ◽

Vol 12 (12) ◽

pp. 1345

Author(s):

Rosilainy Surubi Fernandes ◽

Mariana Rocha David ◽

Filipe Vieira Santos De Abreu ◽

Anielly Ferreira-de-Brito ◽

Noemi R. Gardinali ◽

...

Keyword(s):

Viral Load ◽

Aedes Aegypti ◽

Short Duration ◽

Zika Virus ◽

Rhesus Macaques ◽

Vector Competence ◽

Transmission Dynamics ◽

Zikv Infection ◽

Transmission Rates ◽

Time Points

Despite worldwide efforts to understand the transmission dynamics of Zika virus (ZIKV), scanty evaluation has been made on the vector competence of Aedes aegypti fed directly on viremic human and non-human primates (NHPs). We blood-fed Ae. aegypti from two districts in Rio de Janeiro on six ZIKV infected pregnant rhesus macaques at several time points, half of which were treated with Sofosbuvir (SOF). Mosquitoes were analyzed for vector competence after 3, 7 and 14 days of incubation. Although viremia extended up to eight days post monkey inoculation, only mosquitoes fed on the day of the peak of viremia, recorded on day two, became infected. The influence of SOF treatment could not be assessed because the drug was administered just after mosquito feeding on day two. The global infection, dissemination and transmission rates were quite low (4.09%, 1.91% and 0.54%, respectively); no mosquito was infected when viremia was below 1.26 × 105 RNA copies/mL. In conclusion, Ae. aegypti vector competence for ZIKV from macaques is low, likely to be due to low viral load and the short duration of ZIKV viremia in primates suitable for infecting susceptible mosquitoes. If ZIKV infection in human and macaques behaves similarly, transmission of the Zika virus in nature is most strongly affected by vector density.

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Zika Virus: New Clinical Syndromes and Its Emergence in the Western Hemisphere

Journal of Virology ◽

10.1128/jvi.00252-16 ◽

2016 ◽

Vol 90 (10) ◽

pp. 4864-4875 ◽

Cited By ~ 279

Author(s):

Helen M. Lazear ◽

Michael S. Diamond

Keyword(s):

Zika Virus ◽

Antiviral Agents ◽

Western Hemisphere ◽

Health Concern ◽

Global Public Health ◽

Public Health Concern ◽

Zikv Infection ◽

Clinical Complications ◽

Clinical Syndromes ◽

Potential Interactions

Zika virus (ZIKV) had remained a relatively obscure flavivirus until a recent series of outbreaks accompanied by unexpectedly severe clinical complications brought this virus into the spotlight as causing an infection of global public health concern. In this review, we discuss the history and epidemiology of ZIKV infection, recent outbreaks in Oceania and the emergence of ZIKV in the Western Hemisphere, newly ascribed complications of ZIKV infection, including Guillain-Barré syndrome and microcephaly, potential interactions between ZIKV and dengue virus, and the prospects for the development of antiviral agents and vaccines.

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Estimating risks of importation and local transmission of Zika virus infection

PeerJ ◽

10.7717/peerj.1904 ◽

2016 ◽

Vol 4 ◽

pp. e1904 ◽

Cited By ~ 33

Author(s):

Kyeongah Nah ◽

Kenji Mizumoto ◽

Yuichiro Miyamatsu ◽

Yohei Yasuda ◽

Ryo Kinoshita ◽

...

Keyword(s):

Zika Virus ◽

Arrival Time ◽

Transportation Network ◽

Inverse Function ◽

Network Data ◽

Mosquito Vector ◽

Analysis Model ◽

Zikv Infection ◽

Imported Case ◽

Local Transmission

Background.An international spread of Zika virus (ZIKV) infection has attracted global attention. ZIKV is conveyed by a mosquito vector,Aedesspecies, which also acts as the vector species of dengue and chikungunya viruses.Methods.Arrival time of ZIKV importation (i.e., the time at which the first imported case was diagnosed) in each imported country was collected from publicly available data sources. Employing a survival analysis model in which the hazard is an inverse function of the effective distance as informed by the airline transportation network data, and using dengue and chikungunya virus transmission data, risks of importation and local transmission were estimated.Results.A total of 78 countries with imported case(s) have been identified, with the arrival time ranging from 1 to 44 weeks since the first ZIKV was identified in Brazil, 2015. Whereas the risk of importation was well explained by the airline transportation network data, the risk of local transmission appeared to be best captured by additionally accounting for the presence of dengue and chikungunya viruses.Discussion.The risk of importation may be high given continued global travel of mildly infected travelers but, considering that the public health concerns over ZIKV infection stems from microcephaly, it is more important to focus on the risk of local and widespread transmission that could involve pregnant women. The predicted risk of local transmission was frequently seen in tropical and subtropical countries with dengue or chikungunya epidemic experience.

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Coupled small molecules target RNA interference and JAK/STAT signaling to reduce Zika virus infection in Aedes aegypti

10.1101/2021.08.23.457417 ◽

2021 ◽

Author(s):

Chasity E. Trammell ◽

Gabriela Ramirez ◽

Irma Sanchez-Vargas ◽

Shirley Luckhart ◽

Rushika Perera ◽

...

Keyword(s):

Aedes Aegypti ◽

Small Molecules ◽

Insulin Signaling ◽

Zika Virus ◽

Disease Incidence ◽

Virus Transmission ◽

Zikv Infection ◽

Small Molecule Drugs ◽

Pathway Activation ◽

Target Rna

The recent global Zika epidemics have revealed the significant threat that mosquito-borne viruses pose. There are currently no effective vaccines or prophylactics to prevent Zika virus (ZIKV) infection. Limiting exposure to infected mosquitoes is best way to reduce disease incidence. Recent studies have focused on targeting mosquito reproduction and immune responses to reduce transmission. In particular, previous work evaluated the effect of insulin signaling on antiviral JAK/STAT and RNAi in vector mosquitoes. In this work, we demonstrate that targeting insulin signaling through the repurposing of small molecule drugs results in the activation of both of these antiviral pathways. Activation of this coordinated response additively reduced ZIKV levels in Aedes aegypti mosquitoes. This effect included a quantitatively greater reduction in salivary gland ZIKV levels relative to single pathway activation, indicating the potential for field delivery of these small molecules to substantially reduce virus transmission.

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