The Validation Research on Interaction Between PRRSV 2b Protein and LGALS1

doi:10.11924/j.issn.1000-6850.casb17090090

Abstract

Abstract: The aim is to study the interaction between porcine reproductive and respiratory syndrome virus (PRRSV) 2b protein and host cell proteins. In this study, we used yeast regression test, GST-pull down assay and co- immunoprecipitation assay to verify the interaction between LGALS1 and PRRSV 2b. The results showed that in the yeast regression test, hybrid yeast strain of LGALS1 and2b gene grew blue yeast colonies on SD/-Trp-Leu-His-Ade/AbA/X-α-gal plates. In the GST-pull down assay, the fusion protein bands were detected by Western blot in the samples reacted with 2b protein and LGALS1. In the co-immunoprecipitation assay, samples reacted with 2b protein and LGALS1 were precipitated with HA monoclonal antibody and the corresponding protein bands were detected by Western blot. This paper demonstrates the interaction between LGALS1 and PRRSV 2b protein and provides a theoretical basis for studying PRRSV infection and proliferation.

References

1 刘莹,王凤雪,温永俊,等. 猪繁殖与呼吸综合征病毒非结构蛋白2的研究进展[J]. 中国预防兽医学报,2015,(02):155-158.
2 田志平,薛江东,马徳慧. 猪繁殖与呼吸综合征病毒E蛋白研究进展[J]. 中国畜牧兽医,2012,39(10):42-44.
3 Wang Q, Li Y, Dong H, et al. Identification of host cellular proteins that interact with the M protein of a highly pathogenic porcine reproductive and respiratory syndrome virus vaccine strain[J]. Virol J. 2017,14(1):356-272.
4 Zhang K, Hou Q, Zhong Z, et al. Porcine reproductive and respiratory syndrome virus activates inflammasomes of porcine alveolar macrophages via its small envelope protein E[J]. Virology. 2013, 442(2):156-162.
5 Zhou Y, Fang L, Wang D, et al. Porcine Reproductive and Respiratory Syndrome Virus Infection Induces Stress Granule Formation Depending on Protein Kinase R-like Endoplasmic Reticulum Kinase(PERK) in MARC-145 Cells[J]. Front Cell Infect Microbiol. 2017,4(7):31-37.
6 Martínez-Bosch N, Fernández-Barrena MG, Moreno M, et al.Galectin-1 drives pancreatic carcinogenesis through stroma remodeling and Hedgehog signaling activation[J]. Cancer Res. 2014, 74(13):3512-3524.
7 Park GB, Chung YH, Kim D. Induction of galectin-1 by TLR-dependent PI3K activation enhances epithelial-mesenchymal transition of metastatic ovarian cancer cells[J]. Oncol Rep. 2017, 37(5):3137-3145.
8 Thiemann S, Baum LG. Galectins and Immune Responses-Just How Do They Do Those Things They Do?[J]. Annu Rev Immunol. 2016, 34:243-264.
9杨达汉,邓守全,王哲,等. 猪肺泡巨噬细胞酵母双杂交cDNA文库的构建与鉴定[J]. 中国兽医学报,2017,37(04):632-636+642.
10 ]李先昆,聂智毅,曾日中. 酵母双杂交技术研究与应用进展[J]. 安徽农业科学,2009,37(07):2867-2869+2926.
11 Martínez-Bosch N, Fernández-Barrena MG, Moreno M, et al.Galectin-1 drives pancreatic carcinogenesis through stroma remodeling and Hedgehog signaling activation[J]. Cancer Res. 2014, 74(13):3512-3524.
12 Valach J, Fik Z Strnad H, et al. Smooth muscle actin-expressing stromal fibroblasts in head and neck squamous cell carcinoma: Increased expression of galectin-1 and induction of poor prognosis factors[J]. Int. J. Cancer. 2012, 131, 2499–2508.
13 Yang N, Zhang W, He T, et al. Suppression of Retinal Neovascularization by Inhibition of Galectin-1 in a Murine Model of Oxygen-Induced Retinopathy[J]. J Ophthalmol. 2017, 119(5):224-232.
14 Huang YJ, Shiau AL, Chen SY, et al. Multivalent structure of galectin-1-nanogold complex serves as potential therapeutics for rheumatoid arthritis by enhancing receptor clustering[J]. Eur. Cells Mater. 2012, 23, 170–181.
15 D’Haene N, Sauvage S, Maris C, et al. VEGFR1 and VEGFR2 involvement in extracellular galectin-1- and galectin-3-induced angiogenesis. PLoS ONE, 2013, 8, e67029.
16 Park GB, Chung YH, Kim D. Induction of galectin-1 by TLR-dependent PI3K activation enhances epithelial-mesenchymal transition of metastatic ovarian cancer cells[J]. Oncol Rep. 2017, 37(5):3137-3145.
17 Nam K, Son SH, Oh S, et al. Binding of galectin-1 to integrin β1 potentiates drug resistance by promoting survivin expression in breast cancer cells[J]. Oncotarget. 2017, 92, 1188–1192.
18 Ose R, Oharaa O, Nagase T. Galectin-1 and galectin-3 mediate protocadherin-24-dependent membrane localization of catenin in colon cancer cell line HCT116[J]. Curr. Chem. Genom. 2012, 6, 18–26.
19 Astorgues-Xerri L, Riveiro ME, Tijeras-Raballand,et al. Unraveling galectin-1 as a novel therapeutic target for cancer[J]. Cancer Treat. Rev. 2014, 40, 307–319.
20 Geiger P, Mayer B, Wiest I,et al. Binding of galectin-1 to breast cancer cells MCF7 induces apoptosis and inhibition of proliferation in vitro in a 2D- and 3D- cell culture model[J]. BMC Cancer. 2016, 16(1):870.
21 Takahashi H(1), Takahashi C, Moreland NJ, et al. Establishment of a robust dengue virus NS3-NS5 binding assay for identification of protein-protein interaction inhibitors[J]. Antiviral Res. 2012, 96(3):305-314.
22 Soellick, T.R, Uhrig J.F. Development of an optimized interaction-mating protocol for large-scale yeast two-hybrid analyses[J]. Genome Biology. 2011, 7(4): 1-18.
24 Fong BA, Gillies AR, Ghazi I, et al. Purification of Escherichia coli RNA polymerase using a self-cleaving elastin-like polypeptide tag[J]. Protein Sci. 2010, 19(6):1243-52.