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Discovery of insect and human dengue virus host factors

Abstract

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.

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Figure 1: Genome-wide RNAi screen for dipteran DVHFs.
Figure 2: Injection of Ae. aegypti mosquitoes with dsRNA targeting a DVHF inhibits dengue virus propagation.
Figure 3: Screen for human DVHFs.
Figure 4: Analysis of DENV RNA accumulation after DVHF knockdown.

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Acknowledgements

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.

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Correspondence to James L. Pearson or Mariano A. Garcia-Blanco.

Additional information

Complete list of hits and dsRNA sequence information are available at the DRSC website (http://www.flyrnai.org).

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-6 with Legends, a Supplementary Discussion, Supplementary Methods, Supplementary Tables 1-4 and Supplementary References. (PDF 2788 kb)

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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

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