Dataset Information

The 125th Lys and 145th Thr Amino Acids in the GTPase Domain of Goose Mx Confer Its Antiviral Activity against the Tembusu Virus.

ABSTRACT: The Tembusu virus (TMUV) is an avian pathogenic flavivirus that causes a highly contagious disease and catastrophic losses to the poultry industry. The myxovirus resistance protein (Mx) of innate immune effectors is a key antiviral “workhorse” of the interferon (IFN) system. Although mammalian Mx resistance against myxovirus and retrovirus was witnessed for decades, whether or not bird Mx has anti-flavivirus activity remains unknown. In this study, we found that the transcription of goose Mx (goMx) was obviously driven by TMUV infection, both in vivo and in vitro, and that the titers and copies of TMUV were significantly reduced by goMx overexpression. In both primary (goose embryo fibroblasts, GEFs) and passaged cells (baby hamster kidney cells, BHK21, and human fetal kidney cells, HEK 293T), it was shown that goMx was mainly located in the cytoplasm, and sporadically distributed in the nucleus. The intracellular localization of this protein is attributed to the predicted bipartite nuclear localization signal (NLS; 30 residues: the 441st–471st amino acids of goMx). Intuitively, it seems that the cells with a higher level of goMx expression tend to have lower TMUV loads in the cytoplasm, as determined by an immunofluorescence assay. To further explore the antiviral determinants, a panel of variants was constructed. Two amino acids at the 125th (Lys) and 145th (Thr) positions in GTP-binding elements, not in the L4 loop (40 residues: the 532nd–572nd amino acids of goMx), were vital for the antiviral function of goMx against TMUV in vitro. These findings will contribute to our understanding of the functional significance of the antiviral system in aquatic birds, and the development of goMx could be a valuable therapeutic agent against TMUV.

SUBMITTER: Chen S

PROVIDER: S-EPMC6070871 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

The 125th Lys and 145th Thr Amino Acids in the GTPase Domain of Goose Mx Confer Its Antiviral Activity against the Tembusu Virus.

Chen Shun S Zeng Miao M Liu Peng P Yang Chao C Wang Mingshu M Jia Renyong R Zhu Dekang D Liu Mafeng M Yang Qiao Q Wu Ying Y Zhao Xinxin X Cheng Anchun A

Viruses 20180706 7

The Tembusu virus (TMUV) is an avian pathogenic flavivirus that causes a highly contagious disease and catastrophic losses to the poultry industry. The myxovirus resistance protein (Mx) of innate immune effectors is a key antiviral “workhorse” of the interferon (IFN) system. Although mammalian Mx resistance against myxovirus and retrovirus was witnessed for decades, whether or not bird Mx has anti-flavivirus activity remains unknown. In this study, we found that the transcription of goose ...[more]

PMID: 29986463

Similar Datasets

Project description:Purpose：Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. PBMCs are important immune cells and effectively respond to IFN treatment. Therefore, the goals of this study are to compare the transcriptional profile of the goose IFN (goIFNα, goIFNγ and goIFNλ)-mediated immune response in PBMCs. Methods：Peripheral blood mononuclear cells (PBMCs) were stimulated with recombinant goIFN-α, γ and λ for 3 h, and those treated with cell lysates from pcDNA3.1 (+)-transfected BHK-21 cells were considered as control group. Cells were harvested with 1 mL RNAiso Plus reagent for RNA extraction. Obtained RNA quantity and quality were evaluated using NanoDrop, Qubit and Agilent 2100 Bioanalyzer. After qualification and quantification using an Agilent 2100 Bioanalyzer and ABI Step One Plus Real Time PCR System, the libraries were sequenced with Illumina HiSeqTM 2000. Results：Comparisons between IFN treatment groups and the control group were performed, and DEGs were analysed using the DESeq R package, a model based on the negative binomial distribution. For the statistical analysis, all read counts were normalized by calculating the FPKM value, and further, the FPKM + 1 values were log2 transformed and the means of expression (in log2 FPKMs) were used for further analysis. An P value < 0.05, which was adjusted by the false discovery rate (FDR), was defined as the threshold of DEGs, and log2-fold change ＞ 0 (or < 0) was defined as up-regulated (down-regulated genes). According to the results of DEG analysis, we selected 17 IFN‑responsive genes from the RNA-seq data for validation by RT-qPCR. Conclusion：Our study represents the first detailed analysis of the transcriptional profile of the goose IFN (goIFNα, goIFNγ and goIFNλ) immune response in PBMCs. The stimulation of goIFNs initiated a series of signalling cascades at early stages, leading to the transcriptional regulation of hundreds of DEGs, and each IFN induced a unique and partially overlapping set of ISGs. Most importantly, we identified approximately 101 immune-related genes, which were mainly involved in the toll-like receptor signalling pathway, JAK-STAT signalling pathway and antiviral pathway. Moreover, these results were verified by RT-qPCR, and the correlation analysis of RNA-Seq data and qRT-PCR results showed that some relevant relationships between candidate genes may be related to the balance of the host immune response, which remains to be further studied in the future.

Dataset Information

The 125th Lys and 145th Thr Amino Acids in the GTPase Domain of Goose Mx Confer Its Antiviral Activity against the Tembusu Virus.

Publications

The 125th Lys and 145th Thr Amino Acids in the GTPase Domain of Goose Mx Confer Its Antiviral Activity against the Tembusu Virus.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets