Abstract
Foot-and-mouth disease virus (FMDV) protein 2C is one of the most highly conserved viral proteins among the serotypes of FMDV. However, its effect on host cell response is not very clear. In our previous report, we showed that FMDV protein 2C interacts with cellular protein N-myc and STAT interactor (Nmi), inducing moderate apoptosis in cells. Here, we show that transfection of HEK293T cells with pEGFP-N1-2C or pEGFP-N1-Nmi induces activation of type I interferon promoters, leading to delayed vesicular stomatitis virus (VSV) growth. Using immunoprecipitation and confocal microscopy assays, we found that interferon-induced protein IFP35 interacts with Nmi. Knockdown of IFP35 expression by siRNA abolished pEGFP-N1-2C and pEGFP-N1-Nmi-induced activation of type I interferon promoters and restored VSV growth, suggesting that IFP35 plays a critical role in the type I interferon response induced by FMDV protein 2C. These findings may help to further understand cell responses to FMDV infection.
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Abbreviations
- FMDV:
-
Foot-and-mouth disease virus
- IFP35:
-
Interferon-induced 35-kDa protein
- Nmi:
-
N-myc and STAT interactor
- RNAi:
-
RNA interference
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Acknowledgments
We thank Dr. Jue Liu for his generous assistance. This work was supported by grants from the National Natural Science Foundation of China (#31072117 to SJZ and 31272543 to SJZ)
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705_2014_2147_MOESM1_ESM.tif
Supplementary Fig. 1. Activation of IFN-α promoter by 2C, Nmi or 2C/Nmi. 293T cells (1 × 105) were transfected with the indicated expression plasmids (0.5 mg each) together with the indicated reporter plasmids (0.1 mg each) and the pRL-TK Renilla reporter plasmids (0.01 mg). Reporter assays were performed using a dual-specific luciferase assay kit at 6, 12 and 24 h after the transfection. (TIFF 280 kb)
705_2014_2147_MOESM2_ESM.tif
Supplementary Fig. 2. siRNA resistant form of IFP35 restored type I interferon response in IFP35 RNAi-treated cells. A. Gene sequence of IFP35. The domain targeted by siRNA is highlighted in red. B. Gene sequence of the RNAi-resistant form of IFP35. The siRNA targeting site in (A) was mutated and is highlighted in green. The RNAi resistant form of IFP35 was used to construct pEGFP-mutant vector. C. HEK293T cells were transfected with IFP35 RNAi plasmid or control vector. The IFP35 RNAi stable cell line was selected with puromycin (1 μg/mL in culture medium) and transfected with pEGFP-RNAi-resistant mutant IFP35 vector plus pEGFP empty vector, or pEGFP-N1-2C or pEGFP-N1-Nmi constructs. Twenty-four hours after transfection with pEGFP-N1-2C or pEGFP-N1-Nmi or empty vector as control together with 0.1 μg of IFN-α promoter gene plasmids and 0.01 μg of pRL-TK Renilla reporter plasmids, cells were collected, and the activity of the IFN-α promoter was measured by luciferase reporter gene assay using a dual-specific luciferase assay kit. Data are presented as mean +/- SD, n = 3. ** stands for p < 0.01 and * for p < 0.05. (TIFF 1074 kb)
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Zheng, W., Li, X., Wang, J. et al. A critical role of interferon-induced protein IFP35 in the type I interferon response in cells induced by foot-and-mouth disease virus (FMDV) protein 2C. Arch Virol 159, 2925–2935 (2014). https://doi.org/10.1007/s00705-014-2147-7
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DOI: https://doi.org/10.1007/s00705-014-2147-7