Hepatitis C virus NS5A protein interacts with 2′,5′-oligoadenylate synthetase and inhibits antiviral activity of IFN in an IFN sensitivity-determining region-independent manner

被引:88
作者
Taguchi, T
Nagano-Fujii, M
Akutsu, M
Kadoya, H
Ohgimoto, S
Ishido, S
Hotta, H [1 ]
机构
[1] Kobe Univ, Grad Sch Med, Div Microbiol, Kobe, Hyogo 6500017, Japan
[2] Kobe Univ, Grad Sch Med, Div Diabet Digst & Kidney Dis, Kobe, Hyogo 6500017, Japan
关键词
D O I
10.1099/vir.0.19513-0
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The non-structural protein 5A (NS5A) of hepatitis C virus (HCV) has been implicated in inhibition of antiviral activity of IFN. While previous studies have suggested an interaction between NS5A and the double-stranded RNA-dependent protein kinase (PKR), the possibility still remains that interaction with another molecule(s) is involved in the NS5A-mediated inhibition of IFN. In the present study, we investigated a possible interaction between NS5A and 2',5'-oligoadenylate synthetase (2-5AS), another key molecule in antiviral activity. We observed that NS5A physically interacted with 2-5AS in cultured cells, with an N-terminal portion of NS5A [aa 1-148; NS5A(1-148)] and two separate portions of 2-5AS (aa 52-104 and 184-275) being involved in the interaction. Single point mutations at residue 37 of NS5A affected the degree of the interaction with 2-5AS, with a Phe-to-Leu mutation (F37L) augmenting and a Phe-to-Asn mutation (F37N) diminishing it. Virus rescue assay revealed that the full-length NS5A (NS5A-F) and NS5A(1-148), the latter of which contains neither the IFN sensitivity-determining region (ISDR) nor the PKR-binding domain, significantly counteracted the antiviral activity of IFN. Introduction of a F37N mutation into NS5A(1-148) impaired the otherwise more significant IFN-inhibitory activity of NS5A(1-148). It was also found that the F37N mutation was highly disadvantageous for the replication of an HCV RNA replicon. Taken together, our results suggest the possibility that NS5A interacts with 2-5AS and inhibits the antiviral activity of IFN in an ISDR-independent manner.
引用
收藏
页码:959 / 969
页数:11
相关论文
共 52 条
  • [1] Efficient initiation of HCV RNA replication in cell culture
    Blight, KJ
    Kolykhalov, AA
    Rice, CM
    [J]. SCIENCE, 2000, 290 (5498) : 1972 - 1974
  • [2] An amino-terminal amphipathic α-helix mediates membrane association of the hepatitis C virus nonstructural protein 5A
    Brass, V
    Bieck, E
    Montserret, R
    Wölk, B
    Hellings, JA
    Blum, HE
    Penin, F
    Moradpour, D
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (10) : 8130 - 8139
  • [3] Hepatitis C virus genotypes and risk of hepatocellular carcinoma in cirrhosis: A prospective study
    Bruno, S
    Silini, E
    Crosignani, A
    Borzio, F
    Leandro, G
    Bono, F
    Asti, M
    Rossi, S
    Larghi, A
    Cerino, A
    Podda, M
    Mondelli, MU
    [J]. HEPATOLOGY, 1997, 25 (03) : 754 - 758
  • [4] CHARACTERIZATION OF INSULIN-DEPENDENT DIABETES-MELLITUS INDUCED BY A NEW VARIANT (DK-27) OF ENCEPHALOMYOCARDITIS VIRUS IN DBA/2 MICE
    DAN, K
    SETO, Y
    FUJITA, T
    ASABA, Y
    TAKEI, I
    FUJITA, H
    KATO, R
    [J]. EXPERIMENTAL ANIMALS, 1995, 44 (03): : 211 - 218
  • [5] Hepatitis C virus (HCV) subtype prevalence in Chiang Mai, Thailand, and identification of novel subtypes of HCV major type 6
    Doi, H
    Apichartpiyakul, C
    Ohba, K
    Mizokami, M
    Hotta, H
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 1996, 34 (03) : 569 - 574
  • [6] Sequence analysis of the NS5A protein of European hepatitis C virus 1b isolates and relation to interferon sensitivity
    Duverlie, G
    Khorsi, H
    Castelain, S
    Jaillon, O
    Izopet, J
    Lunel, F
    Eb, F
    Penin, F
    Wychowski, C
    [J]. JOURNAL OF GENERAL VIROLOGY, 1998, 79 : 1373 - 1381
  • [7] Mutations in the nonstructural protein 5A gene and response to interferon in patients with chronic hepatitis C virus 1b infection
    Enomoto, N
    Sakuma, I
    Asahina, Y
    Kurosaki, M
    Murakami, T
    Yamamoto, C
    Ogura, Y
    Izumi, N
    Marumo, F
    Sato, C
    [J]. NEW ENGLAND JOURNAL OF MEDICINE, 1996, 334 (02) : 77 - 81
  • [8] Expression of hepatitis C virus proteins interferes with the antiviral action of interferon independently of PKR-mediated control of protein synthesis
    François, C
    Duverlie, G
    Rebouillat, D
    Khorsi, H
    Castelain, S
    Blum, HE
    Gatignol, A
    Wychowski, C
    Moradpour, D
    Meurs, EF
    [J]. JOURNAL OF VIROLOGY, 2000, 74 (12) : 5587 - 5596
  • [9] Interferon-α inhibits hepatitis C virus subgenomic RNA replication by an MxA-independent pathway
    Frese, M
    Pietschmann, T
    Moradpour, D
    Haller, O
    Bartenschlager, R
    [J]. JOURNAL OF GENERAL VIROLOGY, 2001, 82 : 723 - 733
  • [10] Control of PKR protein kinase by hepatitis C virus nonstructural 5A protein: Molecular mechanisms of kinase regulation
    Gale, M
    Blakely, CM
    Kwieciszewski, B
    Tan, SL
    Dossett, M
    Tang, NM
    Korth, MJ
    Polyak, SJ
    Gretch, DR
    Katze, MG
    [J]. MOLECULAR AND CELLULAR BIOLOGY, 1998, 18 (09) : 5208 - 5218