c-Src protein kinase inhibitors block assembly and maturation of dengue virus
- Department of Microbiology and Molecular Genetics, Harvard Medical School, 200 Longwood Avenue, Boston, MA 02115
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Communicated by John J. Mekalanos, Harvard Medical School, Boston, MA, December 29, 2006 (received for review December 19, 2006)
Abstract
Dengue virus is a mosquito-borne flavivirus that represents an important emerging infectious disease and is an international health concern. Currently, there is no vaccine or effective antiviral therapy to prevent or to treat dengue virus infection. The slow progress in developing antiviral agents might be alleviated by the availability of efficient high-throughput anti-dengue virus screening assays. In this study, we report an immunofluorescence image-based assay suitable for identification of small molecule inhibitors of dengue virus infection and replication. Using this assay, we have discovered that inhibitors of the c-Src protein kinase exhibit a potent inhibitory effect on dengue virus (serotypes 1–4) and murine flavivirus Modoc. Mechanism of action studies demonstrated that the c-Src protein kinase inhibitor dasatinib prevents the assembly of dengue virions within the virus-induced membranous replication complex. These results demonstrate that this cell-based screen may provide a powerful means to identify new potential targets for anti-dengue drug development while simultaneously providing pharmacological probes to investigate dengue virus–host cell interactions at the biochemical level. Given the simplicity and excellent reproducibility of the assay, it should be useful in high-throughput screens of both small molecule and RNAi libraries when implemented on a robotic image-based high-throughput screen (HTS) platform. Given the reasonable clinical safety of inhibitors such as dasatinib and AZD0530, inhibitors of c-Src protein kinase may have the potential to become a new class of anti-dengue viral therapeutic agents.
Footnotes
- *To whom correspondence should be addressed. E-mail: priscilla_yang{at}hms.harvard.edu
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Author contributions: P.L.Y. designed research; J.J.H.C. performed research; J.J.H.C. and P.L.Y. analyzed data; and J.J.H.C. and P.L.Y. wrote the paper.
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The authors declare no conflict of interest.
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This article contains supporting information online at www.pnas.org/cgi/content/full/0611681104/DC1.
- Abbreviations:
- DENV,
- Dengue virus;
- MPA,
- mycophenolic acid;
- moi,
- multiplicity of infection.
- © 2007 by The National Academy of Sciences of the USA
