title: The NS1 gene contributes to the virulence of H5N1 avian influenza viruses.
abstract: In the present study, we explored the genetic basis underlying the virulence and host range of two H5N1 influenza viruses in chickens. A/goose/Guangdong/1/96 (GS/GD/1/96) is a highly pathogenic virus for chickens, whereas A/goose/Guangdong/2/96 (GS/GD/2/96) is unable to replicate in chickens. These two H5N1 viruses differ in sequence by only five amino acids mapping to the PA, NP, M1, and NS1 genes. We used reverse genetics to create four single-gene recombinants that contained one of the sequence-differing genes from nonpathogenic GS/GD/2/96 and the remaining seven gene segments from highly pathogenic GS/GD/1/96. We determined that the NS1 gene of GS/GD/2/96 inhibited the replication of GS/GD/1/96 in chickens, while the substitution of the PA, NP, or M gene did not change the highly pathogenic properties of GS/GD/1/96. Conversely, of the recombinant viruses generated in the GS/GD/2/96 background, only the virus containing the NS1 gene of GS/GD/1/96 was able to replicate and cause disease and death in chickens. The single-amino-acid difference in the sequence of these two NS1 genes resides at position 149. We demonstrate that a recombinant virus expressing the GS/GD/1/96 NS1 protein with Ala149 is able to antagonize the induction of interferon protein levels in chicken embryo fibroblasts (CEFs), but a recombinant virus carrying a Val149 substitution is not capable of the same effect. These results indicate that the NS1 gene is critical for the pathogenicity of avian influenza virus in chickens and that the amino acid residue Ala149 correlates with the ability of these viruses to antagonize interferon induction in CEFs.
authors.affiliation: Harbin Veterinary Research Institute, CAAS, 427 Maduan Street, Harbin 150001, People's Republic of China.
authors.forename: Zejun
authors.initials: Z
authors.lastname: Li
authors.forename: Yongping
authors.initials: Y
authors.lastname: Jiang
authors.forename: Peirong
authors.initials: P
authors.lastname: Jiao
authors.forename: Aiqin
authors.initials: A
authors.lastname: Wang
authors.forename: Fengju
authors.initials: F
authors.lastname: Zhao
authors.forename: Guobin
authors.initials: G
authors.lastname: Tian
authors.forename: Xijun
authors.initials: X
authors.lastname: Wang
authors.forename: Kangzhen
authors.initials: K
authors.lastname: Yu
authors.forename: Zhigao
authors.initials: Z
authors.lastname: Bu
authors.forename: Hualan
authors.initials: H
authors.lastname: Chen
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D002642:Chick Embryo; D002645:Chickens; D004195:Disease Models, Animal; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity*; D005585:Influenza in Birds / Q000821:virology*; D007372:Interferons / Q000037:antagonists & inhibitors; D020125:Mutation, Missense; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000235:genetics / Q000502:physiology; D016601:RNA-Binding Proteins / Q000235:genetics / Q000502:physiology; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics / Q000502:physiology; D011995:Recombination, Genetic; D014758:Viral Core Proteins / Q000235:genetics / Q000502:physiology; D014763:Viral Matrix Proteins / Q000235:genetics / Q000502:physiology; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000502:physiology*; D014764:Viral Proteins / Q000235:genetics / Q000502:physiology; D014774:Virulence / Q000235:genetics; D014779:Virus Replication / Q000235:genetics
chemical_list: C085820:INS1 protein, influenza virus; C488936:M1 protein, Influenza A virus; C048842:NP protein, Influenza A virus; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; C061290:PA protein, influenza viruses; D016601:RNA-Binding Proteins; D014758:Viral Core Proteins; D014763:Viral Matrix Proteins; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins; D007372:Interferons; D012324:RNA-Dependent RNA Polymerase