title: Characterization of the amino acid residues of sendai virus C protein that are critically involved in its interferon antagonism and RNA synthesis down-regulation.
abstract: Sendai virus (SeV) encodes two accessory proteins, V and C, in the alternative reading frames in the P gene that are accessed transcriptionally (V) or translationally (C). The C protein is expressed as a nested set of four C-coterminal proteins, C', C, Y1, and Y2, that use different initiation codons. Using HeLa cell lines constitutively expressing the various C proteins, we previously found that the smallest (the 175-residue Y2) of the four C proteins was fully capable of counteracting the antiviral action of interferons (IFNs) and inhibiting viral RNA synthesis and that the C-terminal half of 106 residues was sufficient for both of these inhibitory functions (A. Kato et al., J. Virol. 75:3802-3810, 2001, and A. Kato et al., J. Virol. 76:7114-7124, 2002). Here, we further generated HeLa cell lines expressing the mutated C (Cm) proteins with charged amino acids substituted for alanine residues at either positions 77 and 80; 114 and 115; 139 and 142; 151, 153, and 154; 156; or 173, 175, and 176. We found that only the mutations at positions 151, 153, and 154 abolished IFN antagonism. All the Cm proteins lost the ability to bind with STAT1 under our assay conditions, regardless of their ability to inhibit IFN signaling. On the other hand, the Cm proteins that altered the tyrosine phosphorylation and dephosphorylation of STAT1 and STAT2 always retained IFN antagonism. Thus, the abnormality of phosphorylation or dephosphorylation appeared to be a cause of the IFN antagonism by SeV C. Regarding viral RNA synthesis inhibition, all mutants but the mutant with replacements at positions 114 and 115 greatly reduced the inhibitory activity, indicating that anti-RNA synthesis by the C protein is governed by amino acids scattered across its C-terminal half. Thus, amino acid sequence requirements differ greatly between IFN antagonism and RNA synthesis inhibition. In addition, we confirmed that another SeV accessory protein, V, does not antagonize IFN.
authors.affiliation: Department of Virology 3, National Institute of Infectious Diseases, Musashi-Murayama, Tokyo 208-0011, Japan. akato@nih.go.jp
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kato
authors.forename: Case
authors.initials: C
authors.lastname: Cortese-Grogan
authors.forename: Sue A
authors.initials: SA
authors.lastname: Moyer
authors.forename: Fumihiro
authors.initials: F
authors.lastname: Sugahara
authors.forename: Takemasa
authors.initials: T
authors.lastname: Sakaguchi
authors.forename: Toru
authors.initials: T
authors.lastname: Kubota
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Otsuki
authors.forename: Masayoshi
authors.initials: M
authors.lastname: Kohase
authors.forename: Masato
authors.initials: M
authors.lastname: Tashiro
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Nagai
mesh_terms: D019943:Amino Acid Substitution; D000998:Antiviral Agents; D004268:DNA-Binding Proteins / Q000378:metabolism; D015536:Down-Regulation; D006367:HeLa Cells; D006801:Humans; D016898:Interferon-alpha / Q000037:antagonists & inhibitors / Q000378:metabolism; D016899:Interferon-beta / Q000037:antagonists & inhibitors / Q000378:metabolism; D007372:Interferons / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010766:Phosphorylation; D012367:RNA, Viral / Q000096:biosynthesis* / Q000187:drug effects*; D050794:STAT1 Transcription Factor; D050795:STAT2 Transcription Factor; D029082:Sendai virus / Q000472:pathogenicity*; D015534:Trans-Activators / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology
chemical_list: D000998:Antiviral Agents; D004268:DNA-Binding Proteins; D016898:Interferon-alpha; D012367:RNA, Viral; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; D050795:STAT2 Transcription Factor; C494084:STAT2 protein, human; D015534:Trans-Activators; D014764:Viral Proteins; C051221:nonstructural C protein, Sendai virus; D016899:Interferon-beta; D007372:Interferons