Antiviral RNAi Response in Culex quinquefasciatus-Derived HSU Cells

Culex spp. mosquitoes are important vectors of viruses, such as West Nile virus, Eastern equine encephalitis virus and Rift valley fever virus. However, their interactions with innate antiviral immunity, especially RNA interference (RNAi), are not well known. Most research on RNAi pathways in mosquitoes is focused on the tropical vector mosquito Aedes aegypti. Here, we investigated the production of arbovirus-specific small RNAs in Cx. quinquefasciatus-derived HSU cells. Furthermore, by silencing RNAi-related proteins, we investigated the antiviral role of these proteins for two different arboviruses: Semliki Forest virus (SFV) and Bunyamwera orthobunyavirus (BUNV). Our results showed an expansion of Ago2 and Piwi6 in Cx. quinquefasciatus compared to Ae. aegypti. While silencing Ago2a and Ago2b increased BUNV replication, only Ago2b showed antiviral activity against SFV. Our results suggest differences in the function of Cx. quinquefasciatus and Ae. aegypti RNAi proteins and highlight the virus-specific function of these proteins in Cx. quinquefasciatus.

[1]  A. Kohl,et al.  An Aedes aegypti-Derived Ago2 Knockout Cell Line to Investigate Arbovirus Infections , 2021, Viruses.

[2]  M. Vermeulen,et al.  PIWI proteomics identifies Atari and Pasilla as piRNA biogenesis factors in Aedes mosquitoes. , 2021, Cell reports.

[3]  Patrick T. Dolan,et al.  Control of RNA viruses in mosquito cells through the acquisition of vDNA and endogenous viral elements , 2019, eLife.

[4]  G. Smagghe,et al.  Arboviruses and the Challenge to Establish Systemic and Persistent Infections in Competent Mosquito Vectors: The Interaction With the RNAi Mechanism , 2019, Front. Physiol..

[5]  G. Ebel,et al.  Small RNA responses of Culex mosquitoes and cell lines during acute and persistent virus infection. , 2019, Insect biochemistry and molecular biology.

[6]  P. Stadler,et al.  The RNA workbench 2.0: next generation RNA data analysis , 2019, Nucleic Acids Res..

[7]  Kazutaka Katoh,et al.  MAFFT online service: multiple sequence alignment, interactive sequence choice and visualization , 2017, Briefings Bioinform..

[8]  M. Varjak,et al.  The antiviral piRNA response in mosquitoes? , 2018, The Journal of general virology.

[9]  Z. Adelman,et al.  Antiviral Immunity and Virus-Mediated Antagonism in Disease Vector Mosquitoes. , 2018, Trends in microbiology.

[10]  Binbin Jin,et al.  piRNA Profiling of Dengue Virus Type 2-Infected Asian Tiger Mosquito and Midgut Tissues , 2018, Viruses.

[11]  Isabelle Dietrich,et al.  Spindle-E Acts Antivirally Against Alphaviruses in Mosquito Cells , 2018, Viruses.

[12]  A. Aravin,et al.  piRNA Biogenesis in Drosophila melanogaster. , 2017, Trends in genetics : TIG.

[13]  Samuel H. Lewis,et al.  Pan-arthropod analysis reveals somatic piRNAs as an ancestral defence against transposable elements , 2017, Nature Ecology & Evolution.

[14]  A. Kohl,et al.  Characterization of the Zika virus induced small RNA response in Aedes aegypti cells , 2017, PLoS neglected tropical diseases.

[15]  M. Watson,et al.  Aedes aegypti Piwi4 Is a Noncanonical PIWI Protein Involved in Antiviral Responses , 2017, mSphere.

[16]  Isabelle Dietrich,et al.  RNA Interference Restricts Rift Valley Fever Virus in Multiple Insect Systems , 2017, mSphere.

[17]  E. Girardi,et al.  Histone-derived piRNA biogenesis depends on the ping-pong partners Piwi5 and Ago3 in Aedes aegypti , 2017, Nucleic acids research.

[18]  Isabelle Dietrich,et al.  The Antiviral RNAi Response in Vector and Non-vector Cells against Orthobunyaviruses , 2017, PLoS neglected tropical diseases.

[19]  P. Miesen,et al.  PIWIs Go Viral: Arbovirus-Derived piRNAs in Vector Mosquitoes , 2016, PLoS pathogens.

[20]  M. Vignuzzi,et al.  Virus-derived DNA drives mosquito vector tolerance to arboviral infection , 2016, Nature Communications.

[21]  Samuel H. Lewis,et al.  Duplication and Diversification of Dipteran Argonaute Genes, and the Evolutionary Divergence of Piwi and Aubergine , 2016, Genome biology and evolution.

[22]  A. Ivens,et al.  Small RNA Profiling in Dengue Virus 2-Infected Aedes Mosquito Cells Reveals Viral piRNAs and Novel Host miRNAs , 2016, PLoS neglected tropical diseases.

[23]  E. Girardi,et al.  Distinct sets of PIWI proteins produce arbovirus and transposon-derived piRNAs in Aedes aegypti mosquito cells , 2015, Nucleic acids research.

[24]  R Core Team,et al.  R: A language and environment for statistical computing. , 2014 .

[25]  Christophe Antoniewski,et al.  Computing siRNA and piRNA overlap signatures. , 2014, Methods in molecular biology.

[26]  B. Williams,et al.  The Developmental Transcriptome of the Mosquito Aedes aegypti, an Invasive Species and Major Arbovirus Vector , 2013, G3: Genes, Genomes, Genetics.

[27]  M. Watson,et al.  Knockdown of piRNA pathway proteins results in enhanced Semliki Forest virus production in mosquito cells , 2013, The Journal of general virology.

[28]  M. Watson,et al.  RNA Interference Targets Arbovirus Replication in Culicoides Cells , 2012, Journal of Virology.

[29]  J. Coppee,et al.  Dicer-2- and Piwi-Mediated RNA Interference in Rift Valley Fever Virus-Infected Mosquito Cells , 2012, Journal of Virology.

[30]  A. Kohl,et al.  New Insights into Control of Arbovirus Replication and Spread by Insect RNA Interference Pathways , 2012, Insects.

[31]  N. Vodovar,et al.  Arbovirus-Derived piRNAs Exhibit a Ping-Pong Signature in Mosquito Cells , 2012, PloS one.

[32]  Z. Adelman,et al.  Production of Virus-Derived Ping-Pong-Dependent piRNA-like Small RNAs in the Mosquito Soma , 2012, PLoS pathogens.

[33]  Catalin C. Barbacioru,et al.  Small RNA profiling of Dengue virus-mosquito interactions implicates the PIWI RNA pathway in anti-viral defense , 2011, BMC Microbiology.

[34]  S. Weaver,et al.  Present and future arboviral threats. , 2010, Antiviral research.

[35]  B. Foy,et al.  Comparative genomics of small RNA regulatory pathway components in vector mosquitoes , 2008, BMC Genomics.

[36]  Olivier Gascuel,et al.  PHYML Online: A Web Server for Fast Maximum Likelihood-Based Phylogenetic Inference , 2018 .

[37]  J. Cross,et al.  Establishment of a line of cells derived from ovarian tissue of Clex quinquefasciatus Say. , 1970, Journal of medical entomology.