Journal of General Virology, Vol 79, 437-446, Copyright © 1998 by Society for General Microbiology

ARTICLES

Identification of a major determinant of mouse neurovirulence of dengue virus type 2 using stably cloned genomic-length cDNA

RC Gualano, MJ Pryor, MR Cauchi, PJ Wright and AD Davidson
Department of Microbiology, Monash University, Clayton, Victoria, Australia.

A genomic-length cDNA clone corresponding to the RNA of dengue virus type 2 (DEN-2) New Guinea C strain (NGC) was constructed in a low copy number vector. The cloned cDNA was stably propagated in Escherichia coil and designated pDVWS501. RNA transcripts produced in vitro from the cDNA using T7 RNA polymerase yielded infectious virus (MON501) upon electroporation into BHK-21 cells. When compared with parental NGC virus, MON501 replicated to similar levels in Aedes albopictus C6/36 cells and showed similar neurovirulence in suckling mice. In contrast, a second genomic-length cDNA clone (pDVWS310) used as an intermediate in the construction of pDVWS501 produced virus (MON310) that replicated well in C6/36 cells but was not neurovirulent in mice. MON310 contained the prM and E genes of the non-neurovirulent PUO-218 strain in an NGC background. There were seven amino acid differences between the prM and E proteins of MON310 and MON501. The differences were generally conservative, with the exception of E residue 126, which was Glu in MON310 and Lys in MON501. To examine the role of this residue in mouse neurovirulence, substitutions of Glu --> Lys and Lys --> Glu were made in MON310 and MON501, respectively. The properties of these mutants clearly demonstrated that Lys at E residue 126 is a major determinant of DEN-2 mouse neurovirulence.

This article has been cited by other articles:

				S. Das, M. R. Pingle, J. Munoz-Jordan, M. S. Rundell, S. Rondini, K. Granger, G.-J. J. Chang, E. Kelly, E. G. Spier, D. Larone, et al. Detection and Serotyping of Dengue Virus in Serum Samples by Multiplex Reverse Transcriptase PCR-Ligase Detection Reaction Assay J. Clin. Microbiol., October 1, 2008; 46(10): 3276 - 3284. [Abstract] [Full Text] [PDF]

				H. Kroschewski, S. P. Lim, R. E. Butcher, T. L. Yap, J. Lescar, P. J. Wright, S. G. Vasudevan, and A. D. Davidson Mutagenesis of the Dengue Virus Type 2 NS5 Methyltransferase Domain J. Biol. Chem., July 11, 2008; 283(28): 19410 - 19421. [Abstract] [Full Text] [PDF]

				B. H. Chua, P. Phuektes, S. A. Sanders, P. K. Nicholls, and P. C. McMinn The molecular basis of mouse adaptation by human enterovirus 71 J. Gen. Virol., July 1, 2008; 89(7): 1622 - 1632. [Abstract] [Full Text] [PDF]

				S. Wati, P. Li, C. J. Burrell, and J. M. Carr Dengue Virus (DV) Replication in Monocyte-Derived Macrophages Is Not Affected by Tumor Necrosis Factor Alpha (TNF-{alpha}), and DV Infection Induces Altered Responsiveness to TNF-{alpha} Stimulation J. Virol., September 15, 2007; 81(18): 10161 - 10171. [Abstract] [Full Text] [PDF]

				H. Malet, M.-P. Egloff, B. Selisko, R. E. Butcher, P. J. Wright, M. Roberts, A. Gruez, G. Sulzenbacher, C. Vonrhein, G. Bricogne, et al. Crystal Structure of the RNA Polymerase Domain of the West Nile Virus Non-structural Protein 5 J. Biol. Chem., April 6, 2007; 282(14): 10678 - 10689. [Abstract] [Full Text] [PDF]

				T. J. Chambers, X. Jiang, D. A. Droll, Y. Liang, W. S. M. Wold, and J. Nickells Chimeric Japanese encephalitis virus/dengue 2 virus infectious clone: biological properties, immunogenicity and protection against dengue encephalitis in mice. J. Gen. Virol., November 1, 2006; 87(Pt 11): 3131 - 3140. [Abstract] [Full Text] [PDF]

				E. Lee, P. J. Wright, A. Davidson, and M. Lobigs Virulence attenuation of Dengue virus due to augmented glycosaminoglycan-binding affinity and restriction in extraneural dissemination. J. Gen. Virol., October 1, 2006; 87(Pt 10): 2791 - 2801. [Abstract] [Full Text] [PDF]

				Z. Zhao, T. Date, Y. Li, T. Kato, M. Miyamoto, K. Yasui, and T. Wakita Characterization of the E-138 (Glu/Lys) mutation in Japanese encephalitis virus by using a stable, full-length, infectious cDNA clone J. Gen. Virol., August 1, 2005; 86(8): 2209 - 2220. [Abstract] [Full Text] [PDF]

				M. J. Pryor, L. Azzola, P. J. Wright, and A. D. Davidson Histidine 39 in the dengue virus type 2 M protein has an important role in virus assembly J. Gen. Virol., December 1, 2004; 85(12): 3627 - 3636. [Abstract] [Full Text] [PDF]

				D. Hayasaka, T. S. Gritsun, K. Yoshii, T. Ueki, A. Goto, T. Mizutani, H. Kariwa, T. Iwasaki, E. A. Gould, and I. Takashima Amino acid changes responsible for attenuation of virus neurovirulence in an infectious cDNA clone of the Oshima strain of Tick-borne encephalitis virus J. Gen. Virol., April 1, 2004; 85(4): 1007 - 1018. [Abstract] [Full Text] [PDF]

				T. J. Harvey, W. J. Liu, X. J. Wang, R. Linedale, M. Jacobs, A. Davidson, T. T. T. Le, I. Anraku, A. Suhrbier, P.-Y. Shi, et al. Tetracycline-Inducible Packaging Cell Line for Production of Flavivirus Replicon Particles J. Virol., January 1, 2004; 78(1): 531 - 538. [Abstract] [Full Text] [PDF]

				W. J. Liu, H. B. Chen, and A. A. Khromykh Molecular and Functional Analyses of Kunjin Virus Infectious cDNA Clones Demonstrate the Essential Roles for NS2A in Virus Assembly and for a Nonconservative Residue in NS3 in RNA Replication J. Virol., July 15, 2003; 77(14): 7804 - 7813. [Abstract] [Full Text] [PDF]

				T. J. Chambers, Y. Liang, D. A. Droll, J. J. Schlesinger, A. D. Davidson, P. J. Wright, and X. Jiang Yellow Fever Virus/Dengue-2 Virus and Yellow Fever Virus/Dengue-4 Virus Chimeras: Biological Characterization, Immunogenicity, and Protection against Dengue Encephalitis in the Mouse Model J. Virol., March 15, 2003; 77(6): 3655 - 3668. [Abstract] [Full Text] [PDF]

				T. P. Monath, J. Arroyo, I. Levenbook, Z.-X. Zhang, J. Catalan, K. Draper, and F. Guirakhoo Single Mutation in the Flavivirus Envelope Protein Hinge Region Increases Neurovirulence for Mice and Monkeys but Decreases Viscerotropism for Monkeys: Relevance to Development and Safety Testing of Live, Attenuated Vaccines J. Virol., February 15, 2002; 76(4): 1932 - 1943. [Abstract] [Full Text] [PDF]

				A. E. Matusan, M. J. Pryor, A. D. Davidson, and P. J. Wright Mutagenesis of the Dengue Virus Type 2 NS3 Protein within and outside Helicase Motifs: Effects on Enzyme Activity and Virus Replication J. Virol., October 15, 2001; 75(20): 9633 - 9643. [Abstract] [Full Text] [PDF]

				F. Guirakhoo, J. Arroyo, K. V. Pugachev, C. Miller, Z.-X. Zhang, R. Weltzin, K. Georgakopoulos, J. Catalan, S. Ocran, K. Soike, et al. Construction, Safety, and Immunogenicity in Nonhuman Primates of a Chimeric Yellow Fever-Dengue Virus Tetravalent Vaccine J. Virol., August 15, 2001; 75(16): 7290 - 7304. [Abstract] [Full Text] [PDF]

				R. J. Hurrelbrink and P. C. McMinn Attenuation of Murray Valley Encephalitis Virus by Site-Directed Mutagenesis of the Hinge and Putative Receptor-Binding Regions of the Envelope Protein J. Virol., August 15, 2001; 75(16): 7692 - 7702. [Abstract] [Full Text] [PDF]

				A. E. Matusan, P. G. Kelley, M. J. Pryor, J. C. Whisstock, A. D. Davidson, and P. J. Wright Mutagenesis of the dengue virus type 2 NS3 proteinase and the production of growth-restricted virus J. Gen. Virol., July 1, 2001; 82(7): 1647 - 1656. [Abstract] [Full Text] [PDF]

				J. Arroyo, F. Guirakhoo, S. Fenner, Z.-X. Zhang, T. P. Monath, and T. J. Chambers Molecular Basis for Attenuation of Neurovirulence of a Yellow Fever Virus/Japanese Encephalitis Virus Chimera Vaccine (ChimeriVax-JE) J. Virol., January 15, 2001; 75(2): 934 - 942. [Abstract] [Full Text]

				F. Guirakhoo, R. Weltzin, T. J. Chambers, Z.-X. Zhang, K. Soike, M. Ratterree, J. Arroyo, K. Georgakopoulos, J. Catalan, and T. P. Monath Recombinant Chimeric Yellow Fever-Dengue Type 2 Virus Is Immunogenic and Protective in Nonhuman Primates J. Virol., June 15, 2000; 74(12): 5477 - 5485. [Abstract] [Full Text]

				R. J. Hurrelbrink, A. Nestorowicz, and P. C. McMinn Characterization of infectious Murray Valley encephalitis virus derived from a stably cloned genome-length cDNA J. Gen. Virol., December 1, 1999; 80(12): 3115 - 3125. [Abstract] [Full Text]