Dengue fever is the most frequent arthropod-borne viral disease of humans, with almost half of the world’s population at risk of infection1. The high prevalence, lack of an effective vaccine, and absence of specific treatment conspire to make dengue fever a global public health threat1,2. Given their compact genomes, dengue viruses (DENV-1–4) and other flaviviruses probably require an extensive number of host factors; however, only a limited number of human, and an even smaller number of insect host factors, have been identified3,4,5,6,7,8,9,10. Here we identify insect host factors required for DENV-2 propagation, by carrying out a genome-wide RNA interference screen in Drosophila melanogaster cells using a well-established 22,632 double-stranded RNA library. This screen identified 116 candidate dengue virus host factors (DVHFs). Although some were previously associated with flaviviruses (for example, V-ATPases and α-glucosidases)3,4,5,7,9,10, most of the DVHFs were newly implicated in dengue virus propagation. The dipteran DVHFs had 82 readily recognizable human homologues and, using a targeted short-interfering-RNA screen, we showed that 42 of these are human DVHFs. This indicates notable conservation of required factors between dipteran and human hosts. This work suggests new approaches to control infection in the insect vector and the mammalian host.
Subscribe to Journal
Get full journal access for 1 year
only $3.83 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Kyle, J. L. & Harris, E. Global spread and persistence of dengue. Annu. Rev. Microbiol. 62, 71–92 (2008)
Mackenzie, J. S., Gubler, D. J. & Petersen, L. R. Emerging flaviviruses: the spread and resurgence of Japanese encephalitis, West Nile and dengue viruses. Nature Med. 10, S98–S109 (2004)
Andoh, T. et al. Effect of bafilomycin A1 on the growth of Japanese encephalitis virus in Vero cells. J. Neurovirol. 4, 627–631 (1998)
Chapel, C. et al. Antiviral effect of α-glucosidase inhibitors on viral morphogenesis and binding properties of hepatitis C virus-like particles. J. Gen. Virol. 87, 861–871 (2006)
Courageot, M. P., Frenkiel, M. P., Dos Santos, C. D., Deubel, V. & Despres, P. α-glucosidase inhibitors reduce dengue virus production by affecting the initial steps of virion morphogenesis in the endoplasmic reticulum. J. Virol. 74, 564–572 (2000)
Emara, M. M. & Brinton, M. A. Interaction of TIA-1/TIAR with West Nile and dengue virus products in infected cells interferes with stress granule formation and processing body assembly. Proc. Natl Acad. Sci. USA 104, 9041–9046 (2007)
Heinz, F. X. et al. Structural changes and functional control of the tick-borne encephalitis virus glycoprotein E by the heterodimeric association with protein prM. Virology 198, 109–117 (1994)
Krishnan, M. N. et al. RNA interference screen for human genes associated with West Nile virus infection. Nature 455, 242–245 (2008)
Nawa, M. Effects of bafilomycin A1 on Japanese encephalitis virus in C6/36 mosquito cells. Arch. Virol. 143, 1555–1568 (1998)
Whitby, K. et al. Castanospermine, a potent inhibitor of dengue virus infection in vitro and in vivo . J. Virol. 79, 8698–8706 (2005)
Hubbard, T. J. et al. Ensembl 2007. Nucleic Acids Res. 35, D610–D617 (2007)
Lawson, D. et al. VectorBase: a home for invertebrate vectors of human pathogens. Nucleic Acids Res. 35, D503–D505 (2007)
Nene, V. et al. Genome sequence of Aedes aegypti, a major arbovirus vector. Science 316, 1718–1723 (2007)
Boutros, M. et al. Genome-wide RNAi analysis of growth and viability in Drosophila cells. Science 303, 832–835 (2004)
Umareddy, I. et al. Dengue virus serotype infection specifies the activation of the unfolded protein response. Virol. J. 4, 91 (2007)
Yu, C. Y., Hsu, Y. W., Liao, C. L. & Lin, Y. L. Flavivirus infection activates the XBP1 pathway of the unfolded protein response to cope with endoplasmic reticulum stress. J. Virol. 80, 11868–11880 (2006)
Littleton, J. T. A genomic analysis of membrane trafficking and neurotransmitter release in Drosophila . J. Cell Biol. 150, F77–F82 (2000)
Pelchen-Matthews, A., Raposo, G. & Marsh, M. Endosomes, exosomes and Trojan viruses. Trends Microbiol. 12, 310–316 (2004)
Krishnan, M. N. et al. Rab 5 is required for the cellular entry of dengue and West Nile viruses. J. Virol. 81, 4881–4885 (2007)
Chu, J. J. & Ng, M. L. Infectious entry of West Nile virus occurs through a clathrin-mediated endocytic pathway. J. Virol. 78, 10543–10555 (2004)
Mosso, C., Galvan-Mendoza, I. J., Ludert, J. E. & del Angel, R. M. Endocytic pathway followed by dengue virus to infect the mosquito cell line C6/36 HT. Virology 378, 193–199 (2008)
Xi, Z., Ramirez, J. L. & Dimopoulos, G. The Aedes aegypti toll pathway controls dengue virus infection. PLoS Pathog. 4, e1000098 (2008)
Kapoor, M. et al. Association between NS3 and NS5 proteins of dengue virus type 2 in the putative RNA replicase is linked to differential phosphorylation of NS5. J. Biol. Chem. 270, 19100–19106 (1995)
Pryor, M. J. et al. Nuclear localization of dengue virus nonstructural protein 5 through its importin α/β-recognized nuclear localization sequences is integral to viral infection. Traffic 8, 795–807 (2007)
Uchil, P. D., Kumar, A. V. & Satchidanandam, V. Nuclear localization of flavivirus RNA synthesis in infected cells. J. Virol. 80, 5451–5464 (2006)
Wang, S. H., Syu, W. J. & Hu, S. T. Identification of the homotypic interaction domain of the core protein of dengue virus type 2. J. Gen. Virol. 85, 2307–2314 (2004)
Ahlquist, P. Parallels among positive-strand RNA viruses, reverse-transcribing viruses and double-stranded RNA viruses. Nature Rev. Microbiol. 4, 371–382 (2006)
Dong, Y. et al. Anopheles gambiae immune responses to human and rodent Plasmodium parasite species. PLoS Pathog. 2, e52 (2006)
Franz, A. W. et al. Engineering RNA interference-based resistance to dengue virus type 2 in genetically modified Aedes aegypti. Proc. Natl Acad. Sci. USA 103, 4198–4203 (2006)
Potter, L. R., Abbey-Hosch, S. & Dickey, D. M. Natriuretic peptides, their receptors, and cyclic guanosine monophosphate-dependent signaling functions. Endocr. Rev. 27, 47–72 (2006)
We thank A. de Silva, C. Lambeth, E. Wagner, F. Scholle, P. Florez de Sessions, J. Umbach, S. Braderick, B. Cullen, J. Nevins, M. Marengo, M. Gromeier, R. Wharton, D. Gubler, E. E. Ooi and S. Vasudevan, and members of the Triangle Flavivirology Group for reagents, time and insights. M.A.G.-B. thanks the late R. Shope for his inspiration and mentorship. We thank the Drosophila RNAi Screening Center (Harvard Medical School), which is funded by National Institutes of Health (NIH) grant RO1 GM067761, for expert assistance. We give special thanks to B. Mathey-Prevot for his support, advice and expertise, and for pointing out the important overlap of our screen with others previously published. We acknowledge funding from the NIH (R21-AI64925 and 5U54-AI057157-05S to M.A.G.-B., 1RO1AI076442 to P.L.Y., and 1R01AI061576-01 to G.D.); the American Society of Microbiology (to J.L.R.); and Johns Hopkins Malaria Research Institute (to J.A.S.-N.). M.A.R. is a Karnovsky Fellow. P.L.Y. acknowledges an award from the Giovanni Armenise–Harvard Foundation. We also acknowledge funding from the North Carolina Biotechnology Center, NIH (1SA0RR024572-1 to M.A.G.-B.), Duke Center for RNA Biology, Duke University School of Medicine, Institute of Genome Sciences and Policy, and Duke Comprehensive Cancer Center (5P30-CA14236) in support of the RNAi facility.
Author Contributions All authors contributed to the strategy and implementation of the work.
Complete list of hits and dsRNA sequence information are available at the DRSC website (http://www.flyrnai.org).
This file contains Supplementary Figures 1-6 with Legends, a Supplementary Discussion, Supplementary Methods, Supplementary Tables 1-4 and Supplementary References. (PDF 2788 kb)
About this article
Cite this article
Sessions, O., Barrows, N., Souza-Neto, J. et al. Discovery of insect and human dengue virus host factors. Nature 458, 1047–1050 (2009). https://doi.org/10.1038/nature07967
Homologs of Human Dengue-Resistance Genes, FKBP1B and ATCAY, Confer Antiviral Resistance in Aedes aegypti Mosquitoes
Current Protocols in Molecular Biology (2019)
Yellow Fever: Integrating Current Knowledge with Technological Innovations to Identify Strategies for Controlling a Re-Emerging Virus
Alternative Experimental Models for Studying Influenza Proteins, Host–Virus Interactions and Anti-Influenza Drugs