Construction of an infectious cDNA clone for Omsk hemorrhagic fever virus, and characterization of mutations in NS2A and NS5

• Published in: Virus research Vol. 155; no. 1; pp. 61 - 68
• Main Authors: Yoshii, Kentaro, Igarashi, Manabu, Ito, Kimihito, Kariwa, Hiroaki, Holbrook, Michael R., Takashima, Ikuo
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 2011
• Subjects: Animals; Body Weight; Cells, Cultured; complementary DNA; Cricetinae; Disease Models, Animal; DNA Mutational Analysis; DNA, Complementary - genetics; Encephalitis Viruses, Tick-Borne - genetics; Encephalitis Viruses, Tick-Borne - growth & development; Encephalitis Viruses, Tick-Borne - pathogenicity; Encephalitis Viruses, Tick-Borne - physiology; Encephalitis, Tick-Borne - pathology; Encephalitis, Tick-Borne - virology; Female; Flavivirus; hemorrhage; Hemorrhagic disease; humans; Infectious cDNA; Kinetics; luciferase; Mice; Mice, Inbred C57BL; molecular cloning; Molecular modelling; Molecular Sequence Data; mutagenesis; Mutant Proteins - genetics; Mutant Proteins - metabolism; Mutation; Mutation, Missense; Omsk hemorrhagic fever; Omsk hemorrhagic fever virus; pathogenesis; phenotype; Replication; replicon; RNA; RNA, Viral - genetics; Sequence Analysis, DNA; Survival Analysis; Tick-borne encephalitis; Viral Load; Viral Nonstructural Proteins - genetics; Viral Nonstructural Proteins - metabolism; Viral replication; Virulence; Virus Replication; viruses; C; E; NS; MOI; Infectious cDNA; ALKV; EM; prM; OHF; OHFV; Flavivirus; KFDV; NLS; Omsk hemorrhagic fever; TBE; RdRp; HDV-RZ; rms; Viral replication; LGTV; ts
• ISSN: 0168-1702; 1872-7492
• DOI: 10.1016/j.virusres.2010.08.023

▶ The construction of Omsk hemorrhagic fever virus. ▶ The recombinant virus shows similar biological properties to the parental virus. ▶ NS2A and NS5 mutations affect viral RNA replication. Omsk hemorrhagic fever virus (OHFV) is a member of the tick-borne encephalitis serocomplex of flaviviruses, and causes hemorrhagic disease in humans. In this study, an infectious cDNA of OHFV was constructed to investigate the molecular mechanisms involved in OHFV pathogenesis for the first time. Our cDNA clone was capable of producing infectious virus which is genetically identical to the parental Guriev strain, and the recombinant virus showed similar biological properties to the parental virus including growth kinetics and virulence characteristics. While characterizing the cDNAs, fortuitous mutations at NS2A position 46 and NS5 position 836 were found to affect viral production. By using a viral replicon expressing luciferase, it was shown that both of the mutations produced a defect in RNA replication and that the NS5 mutation induced a temperature-sensitive phenotype, indicating the importance of these residues in RNA replication. This infectious cDNA will be a useful tool to study the replication and pathogenesis of OHFV.