Presentation on theme: "Genetic evolution of H5N1"— Presentation transcript:
1Genetic evolution of H5N1 Jen-Ren Wang, Ph. D.Department of Medical Laboratory Science and Biotechnology,National Cheng Kung UniversityDepartment of Pathology, National Cheng Kung University HospitalNational Health Research Institutes Tainan Virology Laboratory for Diagnosis and ResearchDepartment of Health, Taiwan Centers for Disease Control contract Virology Laboratory
3Webster, et al. N ENGL J MED. 2006. 355:2174-2177.
4Webster G, et al. N ENGL J MED. 2006. 355:2174-2177.
5Genetic reassortment of H5N1 in Asia from 1999 to 2005 Peiris M. J. S. et al., 2007 Clin Microbiol Rev 20(2), 243–267.
6Phylogenetic relationships of the HA and NP genes of influenza A viruses isolated in Indonesia and VietnamHANPSmith GJ, et al. Virology ;350(2):
7Clade I Clade II, subclade I Smith GJ, et al. Virology ;350(2):
8Smith GJ, et al. Virology. 2006. 5;350(2):258-68.
9Genotypes of H5N1 influenza reassortants in Asia during 2003-2005 (Z+V)(Z+G)(Z)(Z)(Z)(Z+G+W)Chen et al., 2006 PNAS 103,
10Emergence and predominance of an H5N1 variant in China since late 2005 Collection dateNo. of FJ-like viruses2005 July–September1/33 (3) October–December72/136 (53)2006 January–March90/113 (80) April–June103/108 (95)Total266/390 (68)Smith, G. J. D. et al. (2006) Proc. Natl. Acad. Sci. USA 103,
11Antigenetic analysis of different H5N1 lineages GY1IDNFJ-likeGY2VTMGD06, QH-like, Mixed/VNM2Smith, G. J. D. et al. (2006) Proc. Natl. Acad. Sci. USA 103,
12Phylogenetic analysis of H5N1 HA gene from Europe, Africa, and Asia Salzberg SL, et al. EID (5):
14H5N1 pathology/disease Cell tropism: HA, NA Host immune response: NS High viral replication: PA, PB1, PB2, NP
15HA cleavability determinates the tissue tropism Horimoto T, et al. Nature Reviews of Microbiology :
16Effect of HA and NA on replication of influenza virus Influenza viruses bind to sialic acid present on cell surface through the receptor-binding site in the HA molecules followed by receptor-mediated endocytosis during viral entry (Hanson et al., Virology 1992).The NA cleaves the Neu5Ac from the HA molecule to release the progeny virus from the cell membrane and to prevent aggregation of progeny virions (Rogers et al., Virology 1989).This NA enzymatic activity, however, also cleaves the receptor from the target cells.Therefore, the balance between the receptor-binding activity of the HA and the neuraminidase activity of the NA is critical for efficient virus replication in host cells (Kaverin et al., Virology 1998).
17The NA stalk was correlated with the efficiency of virus replication Castrucci MR, et al. Journal of Virology :
18Influenza A virus NS1 protein RNA-binding domain(1-73)Effector domain(73-237)RNA-binding domain (19-38)Nuclear localization signal (34-38, )Inhibition of PKR activation and regulation of vRNA synthesis ( )Nuclear export signal ( )30 kDa subunit of Cleavage and polyadenylation specific factor (CPSF) binding site (186)poly-A binding protein II (PABII) binding domain ( )PDZ domain ligand ( )
19NS1 as IFN-α/β antagonist Influenza A virusIRF-3/IRF-7+P300 and CREB-binding proteindsRNANS1cytoplsamIFN/ISREAAUAAAAntiviral pre-mRNAs5’ capAAAAA………nucleusCPSF30kPABIIXsplicingsnRNA U6PKRRIG-INemeroff M. et al., Mol.Cell., 1998Yuan L. et al., Virology, 1995Zhongying C. et al., EMBO, 1999Yun Q. et al., J.Virol., 1994Mibayashi M et al., J. Virol., 2007NS1 binds to dsRNA and RIG-I as IFN-α/β antagonistNS1 binds to snRNA U6, poly-A mRNA, CPSF, PABII and PKR against antiviral genes expression
20PDZ ligand motif in NS1 as a potential virulence determinant Large-scale sequence analysis of avian influenza isolatesObenauer et al. science. 20061997PDZ domainRegulating the activity and trafficking of membrane proteinsMaintaining cell polarity and morphologyOrganizing postsynaptic density in neuronal cells20031918
21Importance of NS1A D92E mutation in NS1 strongly affect the virulence of influenza virus, eg: the H5N1 avian flu virus (Seo et al. 2004) .C-terminal PDZ domain ligand in NS1 act as a potential virulence determinant (Krug et al. 2006) .Amino acid 89 in the NS1 protein as being critical for binding to p85 (Hale et al. 2006).Phe-103 and Met-106 residues in NS1 is critical for CPSF binding (Kochs et al. 2007).
22NS1 gene E92 and del 80-84 may affect RNA binding affinity H5N1 of Hong Kong outbreakNat.Struct.Mol.Biol. 2006E92 and del may affect RNA binding affinityCytokine-resistance
23Role of PB2 genesA E627K mutation in PB2 strongly affect the virulence of influenza virus, eg: the 1918 flu virus and the H5N1 avian flu virus (Gillis et al. 2005).Majority of avian viruses have PB2 E627 except Qinghai lineage, it may be one adaptation of virus to mammalian host.
25Schematic diagram of chimeric and single amino acid PB2 mutants, with their virulence in mice (MLD50)mutation at position 627 in the PB2 protein influenced the outcome of infection in miceHatta M, et al. Science ;293(5536):