Volume 75, Issue 6
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645


Although much is known about the ecology, epidemiology, and molecular biology of mosquito-borne viruses, the viral factors that allow transmission by mosquitoes to humans or animals remain unknown. Using infectious clones of disseminating (Asibi) and non-disseminating (17D) yellow fever viruses (YFV), we produced chimeric viruses to evaluate the role of different viral genes in dissemination. Previously, we showed that virus produced from an infectious clone containing the structural genes of 17D in Asibi disseminated from the mosquito midgut at a rate of 31%, indicating that some genetic determinants of dissemination must lie within the non-structural (NS) protein genes or 3′ non-coding region (NCR). We chose to investigate the roles of NS2A, NS4B, and the 3′NCR in YFV dissemination. Substitution of the 17D NS2A or NS4B into Asibi significantly attenuated YFV dissemination, demonstrating that this is a multigenic property. There was no difference in dissemination after substitution of the 17D 3′NCR.


Article metrics loading...

The graphs shown below represent data from March 2017
Loading full text...

Full text loading...



  1. Higgs S, 2004. How do mosquito vectors live with their viruses? Gillespie SH, Smith GL, Osbourn A, eds. Microbe-Vector Interactions in Vector-Borne Diseases. Cambridge, United Kingdom: Cambridge University Press, 103–137.
  2. Hombach J, Barrett AD, Cardosa MJ, Deubel V, Guzman M, Kurane I, Roehrig JT, Sabchareon A, Kieny MP, 2005. Review on flavivirus vaccine development. Proceedings of a meeting jointly organized by the World Health Organization and the Thai Ministry of Public Health, 26–27 April 2004, Bangkok, Thailand. Vaccine 23 : 2689–2695. [Google Scholar]
  3. McElroy KL, Tsetsarkin KA, Vanlandingham DL, Higgs S, 2006. Role of the yellow fever virus structural protein genes in viral dissemination from the Aedes aegypti mosquito midgut. J Gen Virol 87 : 2993–3001. [Google Scholar]
  4. Brault AC, Powers AM, Weaver SC, 2002. Vector infection determinants of venezuelan equine encephalitis virus reside within the E2 envelope glycoprotein. J Virol 76 : 6387–6392. [Google Scholar]
  5. Brault AC, Powers AM, Ortiz D, Estrada-Franco JG, Navarro-Lopes R, Weaver SC, 2004. Venezuelan equine encephalitis emergence: enhanced vector infection from a single amino acid substitution in the envelope protein. Proc Natl Acad Sci U S A 101 : 11344–11349. [Google Scholar]
  6. Ludwig GV, Isreal BA, Christensen BM, Yuill TM, Schultz KT, 1991. Role of La Crosse virus glycoproteins in attachment of virus to host cells. Virology 181 : 564–571. [Google Scholar]
  7. Myles KM, Pierro DJ, Olson KE, 2003. Deletions in the putative cell receptor-binding domain of Sindbis virus strain MRE16 E2 glycoprotein reduce midgut infectivity in Aedes aegypti. J Virol 77 : 8872–8881. [Google Scholar]
  8. Sundin DR, Beaty BJ, Nathanson M, Gonzales-Scarano F, 1987. A G1 glycoprotein epitope of La Crosse virus: a determinant of infection of Aedes triseriatus. Science 235 : 591–593. [Google Scholar]
  9. Woodward TM, Miller BR, Beaty BJ, Trent DW, Roehrig JT, 1991. A single amino acid change in the E2 glycoprotein of Venezuelan equine encephalitis virus affects replication and dissemination in Aedes aegypti mosquitoes. J Gen Virol 72 : 2431–2435. [Google Scholar]
  10. Johnson BW, Chambers TV, Crabtree MB, Bhatt TR, Guirakhoo F, Monath TP, Miller BR, 2002. Growth characteristics of ChimeriVax-DEN2 vaccine virus in Aedes aegypti and Aedes albopictus mosquitoes. Am J Trop Med Hyg 67 : 260–265. [Google Scholar]
  11. Johnson BW, Chambers TV, Crabtree MB, Arroyo J, Monath TP, Miller BR, 2003. Growth characteristics of the veterinary vaccine candidate ChimeriVax-West Nile (WN) virus in Aedes and Culex mosquitoes. Med Vet Entomol 17 : 235–243. [Google Scholar]
  12. Troyer JM, Hanley KA, Whitehead SS, Strickman D, Karron RA, Durbin AP, Murphy BR, 2001. A live attenuated recombinant dengue-4 virus vaccine candidate with restricted capacity for dissemination in mosquitoes and lack of transmission from vaccinees to mosquitoes. Am J Trop Med Hyg 65 : 414–419. [Google Scholar]
  13. Whitehead SS, Hanley KA, Blaney JA, Gilmore LE, Elkins WR, Murphy BR, 2003. Substitution of the structural genes of dengue virus type 4 with those of type 2 results in chimeric vaccine candidates which are attenuated in mosquitoes, mice, and rhesus monkeys. Vaccine 21 : 4307–4316. [Google Scholar]
  14. Jennings AD, Gibson CA, Miller BR, Mathews JH, Mitchell CJ, Roehrig JT, Wood DJ, Taffs F, Sil BK, Whitby SN, Whitby JE, Monath TP, Minor PD, Sanders PG, Barrett ADT, 1994. Analysis of a yellow fever virus isolated from a fatal case of vaccine-associated human encephalitis. J Infect Dis 169 : 512–518. [Google Scholar]
  15. Whitman L, 1939. Failure of Aedes aegypti to transmit yellow fever cultured virus (17D). Am J Trop Med Hyg 19 : 19–26. [Google Scholar]
  16. Miller BR, Adkins D, 1988. Biological characterization of plaque-size variants of yellow fever virus in mosquitoes and mice. Acta Virol 32 : 227–234. [Google Scholar]
  17. McElroy KL, Tsetsarkin KA, Vanlandingham DL, Higgs S, 2005. Characterization of an infectious clone of the wild-type yellow fever virus Asibi strain that is able to infect and disseminate in mosquitoes. J Gen Virol 86 : 1747–1751. [Google Scholar]
  18. Bredenbeek PJ, Kooi EA, Lindenbach B, Huijkman N, Rice CM, Spaan WJ, 2003. A stable full-length yellow fever virus cDNA clone and the role of conserved RNA elements in flavivirus replication. J Gen Virol 84 : 1261–1268. [Google Scholar]
  19. Kummerer BM, Rice CM, 2002. Mutations in the yellow fever virus nonstructural protein NS2A selectively block production of infectious particles. J Virol 76 : 4773–4784. [Google Scholar]
  20. Mackenzie JM, Kromykh AA, Jones MK, Westaway EG, 1998. Subcellular localization and some biochemical properties of the flavivirus Kunjin nonstructural proteins NS2A and NS4A. Virology 245 : 203–215. [Google Scholar]
  21. Bennett SN, Holmes EC, Chirivella M, Rodriguez DM, Beltran M, Vorndam V, Gubler DJ, McMillan WO, 2003. Seletion-driven evolution of emergent dengue virus. Mol Biol Evol 20 : 1650–1658. [Google Scholar]
  22. Dunster LM, Wang H, Ryman KD, Miller BR, Watowich SJ, Minor PD, Barrett ADT, 1991. Molecular and biological changes associated with HeLa cell attenuation of wild-type yellow fever virus. Virology 261 : 309–318. [Google Scholar]
  23. Wang H, Ryman KD, Jennings AD, Wood AJ, Taffs F, Minor PD, Sanders PG, Barrett AD, 1995. Comparison of the genomes of the wild-type French viscerotropic strain of yellow fever virus with its vaccine derivative French neurotropic vaccine. J Gen Virol 76 : 2749–2755. [Google Scholar]
  24. Proutski V, Gaunt MW, Gould EA, Holmes EC, 1997. Secondary structure of the 3′-untranslated region of yellow fever virus: implications for virulence, attenuation and vaccine development. J Gen Virol 78 : 1543–1549. [Google Scholar]
  25. Vanlandingham DL, Hong C, Klingler K, Tsetsarkin K, McElroy KL, Powers AM, Lehane MJ, Higgs S, 2005. Differential infectivities of o’nyong-nyong and chikungunya virus isolates in Anopheles gambiae and Aedes aegypti mosquitoes. Am J Trop Med Hyg 72 : 616–621. [Google Scholar]
  26. Vanlandingham DL, Tsetsarkin KA, Klingler KA, Hong C, McElroy KL, Lehane MJ, Higgs S, 2006. Determinants of vector specificity of o’nyong-nyong and chikungunya virus isolates in Anopheles gambiae and Aedes aegypti mosquitoes. Am J Trop Med Hyg 74 : 663–669. [Google Scholar]
  27. Blaney JA, Johnson DH, Manipon GG, Firestone CY, Hanson CT, Murphy BR, Whitehead SS, 2002. Genetic basis of attenuation of dengue virus type 4 small plaque mutants with restricted replication in suckling mice and in SCID mice transplanted with human liver cells. Virology 300 : 125–139. [Google Scholar]
  28. Romoser WS, Wasielowski LP, Pushko P, Kondig JP, Lerdthusnee K, Neira M, Ludwig GV, 2004. Evidence for arbovirus dissemination conduits from the mosquito (Diptera: Culicidae) midgut. J Med Entomol 41 : 467–475. [Google Scholar]
  29. Blaney JA, Hanson CM, Firestone CY, Hanley KA, Murphy BR, Whitehead SS, 2004. Genetically modified, live attenuated dengue virus type 3 vaccine candidates. Am J Trop Med Hyg 71 : 811–821. [Google Scholar]
  30. Hanley KA, Manlucu LR, Gilmore LE, Blaney JE, Hanson CT, Murphy BR, Whitehead SS, 2003. A trade-off in replication in mosquito versus mammalian systems conferred by a point mutation in the NS4B protein of dengue virus type 4. Virology 312 : 222–232. [Google Scholar]
  31. Muñoz-Jordán JL, Sánchez-Burgos GG, Laurent-Rolle M, García-Sastre A, 2003. Inhibition of interferon signaling by dengue virus. Proc Natl Acad Sci U S A 100 : 14333–14338. [Google Scholar]
  32. Muñoz-Jordán JL, Laurent-Rolle M, Ashour J, Martínez-Sobrido L, Ashok M, Lipkin WI, García-Sastre A, 2005. Inhibition of alpha/beta interferon signaling by the NS4B protein of flaviviruses. J Virol 79 : 8004–8013. [Google Scholar]

Data & Media loading...

  • Received : 28 Jun 2006
  • Accepted : 28 Jul 2006

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error