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Characterization of dengue virus 3’UTR RNA binding proteins in mosquitoes reveals that AeStaufen reduces subgenomic flaviviral RNA in saliva

Dengue viruses (DENV) are expanding global pathogens that are transmitted through the bite of mosquitoes, mostly Aedes aegypti. As RNA viruses, DENV rely on RNA-binding proteins (RBPs) to complete their life cycle. Alternatively, RBPs can act as restriction factors that prevent DENV multiplication....

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Autores principales: Yeh, Shih-Chia, Diosa-Toro, Mayra, Tan, Wei-Lian, Rachenne, Florian, Hain, Arthur, Yeo, Celestia Pei Xuan, Bribes, Inès, Xiang, Benjamin Wong Wei, Sathiamoorthy Kannan, Gayathiri, Manuel, Menchie Casayuran, Missé, Dorothée, Mok, Yu Keung, Pompon, Julien
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9531803/
https://www.ncbi.nlm.nih.gov/pubmed/36121894
http://dx.doi.org/10.1371/journal.ppat.1010427
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author Yeh, Shih-Chia
Diosa-Toro, Mayra
Tan, Wei-Lian
Rachenne, Florian
Hain, Arthur
Yeo, Celestia Pei Xuan
Bribes, Inès
Xiang, Benjamin Wong Wei
Sathiamoorthy Kannan, Gayathiri
Manuel, Menchie Casayuran
Missé, Dorothée
Mok, Yu Keung
Pompon, Julien
author_facet Yeh, Shih-Chia
Diosa-Toro, Mayra
Tan, Wei-Lian
Rachenne, Florian
Hain, Arthur
Yeo, Celestia Pei Xuan
Bribes, Inès
Xiang, Benjamin Wong Wei
Sathiamoorthy Kannan, Gayathiri
Manuel, Menchie Casayuran
Missé, Dorothée
Mok, Yu Keung
Pompon, Julien
author_sort Yeh, Shih-Chia
collection PubMed
description Dengue viruses (DENV) are expanding global pathogens that are transmitted through the bite of mosquitoes, mostly Aedes aegypti. As RNA viruses, DENV rely on RNA-binding proteins (RBPs) to complete their life cycle. Alternatively, RBPs can act as restriction factors that prevent DENV multiplication. While the importance of RBPs is well-supported in humans, there is a dearth of information about their influence on DENV transmission by mosquitoes. Such knowledge could be harnessed to design novel, effective interventions against DENV. Here, we successfully adapted RNA-affinity chromatography coupled with mass spectrometry–a technique initially developed in mammalian cells–to identify RBPs in Ae. aegypti cells. We identified fourteen RBPs interacting with DENV serotype 2 3’UTR, which is involved in the viral multiplication and produces subgenomic flaviviral RNA (sfRNA). We validated the RNA affinity results for two RBPs by confirming that AePur binds the 3’UTR, whereas AeStaufen interacts with both 3’UTR and sfRNA. Using in vivo functional evaluation, we determined that RBPs like AeRan, AeExoRNase, and AeRNase have pro-viral functions, whereas AeGTPase, AeAtu, and AePur have anti-viral functions in mosquitoes. Furthermore, we showed that human and mosquito Pur homologs have a shared affinity to DENV2 RNA, although the anti-viral effect is specific to the mosquito protein. Importantly, we revealed that AeStaufen mediates a reduction of gRNA and sfRNA copies in several mosquito tissues, including the salivary glands and that AeStaufen-mediated sfRNA reduction diminishes the concentration of transmission-enhancing sfRNA in saliva, thereby revealing AeStaufen’s role in DENV transmission. By characterizing the first RBPs that associate with DENV2 3’UTR in mosquitoes, our study unravels new pro- and anti-viral targets for the design of novel therapeutic interventions as well as provides foundation for studying the role of RBPs in virus-vector interactions.
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spelling pubmed-95318032022-10-05 Characterization of dengue virus 3’UTR RNA binding proteins in mosquitoes reveals that AeStaufen reduces subgenomic flaviviral RNA in saliva Yeh, Shih-Chia Diosa-Toro, Mayra Tan, Wei-Lian Rachenne, Florian Hain, Arthur Yeo, Celestia Pei Xuan Bribes, Inès Xiang, Benjamin Wong Wei Sathiamoorthy Kannan, Gayathiri Manuel, Menchie Casayuran Missé, Dorothée Mok, Yu Keung Pompon, Julien PLoS Pathog Research Article Dengue viruses (DENV) are expanding global pathogens that are transmitted through the bite of mosquitoes, mostly Aedes aegypti. As RNA viruses, DENV rely on RNA-binding proteins (RBPs) to complete their life cycle. Alternatively, RBPs can act as restriction factors that prevent DENV multiplication. While the importance of RBPs is well-supported in humans, there is a dearth of information about their influence on DENV transmission by mosquitoes. Such knowledge could be harnessed to design novel, effective interventions against DENV. Here, we successfully adapted RNA-affinity chromatography coupled with mass spectrometry–a technique initially developed in mammalian cells–to identify RBPs in Ae. aegypti cells. We identified fourteen RBPs interacting with DENV serotype 2 3’UTR, which is involved in the viral multiplication and produces subgenomic flaviviral RNA (sfRNA). We validated the RNA affinity results for two RBPs by confirming that AePur binds the 3’UTR, whereas AeStaufen interacts with both 3’UTR and sfRNA. Using in vivo functional evaluation, we determined that RBPs like AeRan, AeExoRNase, and AeRNase have pro-viral functions, whereas AeGTPase, AeAtu, and AePur have anti-viral functions in mosquitoes. Furthermore, we showed that human and mosquito Pur homologs have a shared affinity to DENV2 RNA, although the anti-viral effect is specific to the mosquito protein. Importantly, we revealed that AeStaufen mediates a reduction of gRNA and sfRNA copies in several mosquito tissues, including the salivary glands and that AeStaufen-mediated sfRNA reduction diminishes the concentration of transmission-enhancing sfRNA in saliva, thereby revealing AeStaufen’s role in DENV transmission. By characterizing the first RBPs that associate with DENV2 3’UTR in mosquitoes, our study unravels new pro- and anti-viral targets for the design of novel therapeutic interventions as well as provides foundation for studying the role of RBPs in virus-vector interactions. Public Library of Science 2022-09-19 /pmc/articles/PMC9531803/ /pubmed/36121894 http://dx.doi.org/10.1371/journal.ppat.1010427 Text en © 2022 Yeh et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Yeh, Shih-Chia
Diosa-Toro, Mayra
Tan, Wei-Lian
Rachenne, Florian
Hain, Arthur
Yeo, Celestia Pei Xuan
Bribes, Inès
Xiang, Benjamin Wong Wei
Sathiamoorthy Kannan, Gayathiri
Manuel, Menchie Casayuran
Missé, Dorothée
Mok, Yu Keung
Pompon, Julien
Characterization of dengue virus 3’UTR RNA binding proteins in mosquitoes reveals that AeStaufen reduces subgenomic flaviviral RNA in saliva
title Characterization of dengue virus 3’UTR RNA binding proteins in mosquitoes reveals that AeStaufen reduces subgenomic flaviviral RNA in saliva
title_full Characterization of dengue virus 3’UTR RNA binding proteins in mosquitoes reveals that AeStaufen reduces subgenomic flaviviral RNA in saliva
title_fullStr Characterization of dengue virus 3’UTR RNA binding proteins in mosquitoes reveals that AeStaufen reduces subgenomic flaviviral RNA in saliva
title_full_unstemmed Characterization of dengue virus 3’UTR RNA binding proteins in mosquitoes reveals that AeStaufen reduces subgenomic flaviviral RNA in saliva
title_short Characterization of dengue virus 3’UTR RNA binding proteins in mosquitoes reveals that AeStaufen reduces subgenomic flaviviral RNA in saliva
title_sort characterization of dengue virus 3’utr rna binding proteins in mosquitoes reveals that aestaufen reduces subgenomic flaviviral rna in saliva
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9531803/
https://www.ncbi.nlm.nih.gov/pubmed/36121894
http://dx.doi.org/10.1371/journal.ppat.1010427
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