Drosophila RNAi screen identifies host genes important for influenza virus replication (original) (raw)

Nature volume 454, pages 890–893 (2008)Cite this article

Abstract

All viruses rely on host cell proteins and their associated mechanisms to complete the viral life cycle. Identifying the host molecules that participate in each step of virus replication could provide valuable new targets for antiviral therapy, but this goal may take several decades to achieve with conventional forward genetic screening methods and mammalian cell cultures. Here we describe a novel genome-wide RNA interference (RNAi) screen in Drosophila1 that can be used to identify host genes important for influenza virus replication. After modifying influenza virus to allow infection of Drosophila cells and detection of influenza virus gene expression, we tested an RNAi library against 13,071 genes (90% of the Drosophila genome), identifying over 100 for which suppression in Drosophila cells significantly inhibited or stimulated reporter gene (Renilla luciferase) expression from an influenza-virus-derived vector. The relevance of these findings to influenza virus infection of mammalian cells is illustrated for a subset of the Drosophila genes identified; that is, for three implicated Drosophila genes, the corresponding human homologues ATP6V0D1, COX6A1 and NXF1 are shown to have key functions in the replication of H5N1 and H1N1 influenza A viruses, but not vesicular stomatitis virus or vaccinia virus, in human HEK 293 cells. Thus, we have demonstrated the feasibility of using genome-wide RNAi screens in Drosophila to identify previously unrecognized host proteins that are required for influenza virus replication. This could accelerate the development of new classes of antiviral drugs for chemoprophylaxis and treatment, which are urgently needed given the obstacles to rapid development of an effective vaccine against pandemic influenza and the probable emergence of strains resistant to available drugs.

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Acknowledgements

We thank M. McGregor and K. Wells for technical assistance, M. Hatta, S. Yamada and M. Ito for an H5N1 influenza virus, S. Watanabe and Y. Hatta for FVG-R and FVG-G viruses, R. Kalejta and J. Bruce for adenovirus and murine leukaemia virus vectors, T. Noda for electron microscopy, J. Gilbert for editing the manuscript and Y. Kawaoka for illustrations. This work was supported, in part, by US National Institute of Allergy and Infectious Diseases Public Health Service research grants, by the US National Institute of General Medical Sciences grant GM35072, by a grant-in-aid for Specially Promoted Research and by a contract research fund for the Program of Funding Research Centers for Emerging and Reemerging Infectious Diseases from the Ministry of Education, Culture, Sports, Science and Technology, and by grants-in-aid from the Ministry of Health, Labor, Welfare of Japan. P.A. is a Howard Hughes Medical Institute investigator.

Author Contributions L.H., A.S., P.A. and Y.K. designed research; L.H., A.S., T.W. and C.A.N. performed research; L.H., A.S., T.W., E.S., M.A.N., P.A. and Y.K. analysed data; and L.H., A.S., T.W., P.A. and Y.K. wrote the manuscript.

Author information

Author notes

  1. Akira Sakurai
    Present address: Present address: First Department of Forensic Science, National Research Institute of Police Science, 6-3-1 Kashiwanoha, Kashiwa 277-0882, Japan.,
  2. Linhui Hao and Akira Sakurai: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute for Molecular Virology,,
    Linhui Hao, Ericka Sorensen & Paul Ahlquist
  2. Howard Hughes Medical Institute,,
    Linhui Hao & Paul Ahlquist
  3. Department of Pathobiological Sciences, and,
    Akira Sakurai, Tokiko Watanabe & Yoshihiro Kawaoka
  4. Departments of Statistics and of Biostatistics and Medical Informatics, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA,
    Michael A. Newton
  5. Faculty of Veterinary Medicine, and,,
    Chairul A. Nidom
  6. Collaborating Research Center-Emerging & Reemerging Infectious Diseases, Tropical Disease Centre, Airlangga University, Surabaya 60115, Indonesia ,
    Chairul A. Nidom
  7. Division of Virology, Department of Microbiology and Immunology, and,
    Yoshihiro Kawaoka
  8. International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan ,
    Yoshihiro Kawaoka
  9. Division of Zoonosis, Department of Microbiology and Infectious Diseases, Graduate School of Medicine, Kobe University, Kobe 650-0017, Japan
    Yoshihiro Kawaoka

Authors

  1. Linhui Hao
  2. Akira Sakurai
  3. Tokiko Watanabe
  4. Ericka Sorensen
  5. Chairul A. Nidom
  6. Michael A. Newton
  7. Paul Ahlquist
  8. Yoshihiro Kawaoka

Corresponding authors

Correspondence toPaul Ahlquist or Yoshihiro Kawaoka.

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Hao, L., Sakurai, A., Watanabe, T. et al. Drosophila RNAi screen identifies host genes important for influenza virus replication.Nature 454, 890–893 (2008). https://doi.org/10.1038/nature07151

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Editorial Summary

Flu in a fruit fly: the search for new antivirals

Although several alternative antivirals are available for the treatment of influenza virus infection, the risk that viruses resistant to these drugs will emerge means it is important to continue the search for new antiviral targets. Hao et al. have recruited a new ally to this cause: by modifying influenza virus so that it can infect the cells of the fruit fly Drosophila, they have been able to use a powerful genome-wide RNA interference (RNAi) screen to identify scores of host genes that the pathogen requires for successful infection. Several host proteins were found that have key functions in the replication of H5N1 and H1N1 influenza A viruses — but not other viruses — in human cells. The same strategy should be applicable to other viruses, as long as at least part of their replication cycle can be supported in Drosophila cells.