Carboxyl terminal activating region 3 of latent membrane protein 1 encoded by the Epstein‑Barr virus regulates cell proliferation and protein expression in NP69 cells

Zhang,Zhi‑Wei; Zhang,He‑Liang; Yu,Yan‑Hui; Ouyang,Yong‑Mei; Chen,Zhu‑Chu; He,Xiu‑Sheng; He,Zhi-Min

doi:10.3892/mmr.2019.10859

Carboxyl terminal activating region 3 of latent membrane protein 1 encoded by the Epstein‑Barr virus regulates cell proliferation and protein expression in NP69 cells

Authors:
- Zhi‑Wei Zhang
- He‑Liang Zhang
- Yan‑Hui Yu
- Yong‑Mei Ouyang
- Zhu‑Chu Chen
- Xiu‑Sheng He
- Zhi-Min He
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Affiliations: Cancer Research Institute of Medical College, University of South China, Key Laboratory of Cancer Cellular and Molecular Pathology of Hunan, Hengyang, Hunan 421001, P.R. China, Cancer Research Institute, Central South University, Xiangya School of Medicine, Changsha, Hunan 410078, P.R. China
Published online on: December 4, 2019 https://doi.org/10.3892/mmr.2019.10859
Pages: 720-730
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

In the present study, the mechanism by which carboxyl terminal activating region 3 (CTAR3) of latent membrane protein 1 (LMP1), encoded by the Epstein‑Barr virus, regulated cell proliferation and protein expression was investigated in the nasopharyngeal epithelial cell line NP69. The deletion mutant LMP1 (LMP1Δ232‑351; amino acid residues including 232‑351 codons in CTAR3 deleted) was generated by polymerase chain reaction. An NP69‑LMP1Δ232‑351 cell line was established by retroviral infection. Finally, cell proliferation and protein expression of NP69 cells expressing LMP1Δ232‑351 were examined using a cell growth curve and western blot analysis. The results demonstrated: i) The proliferation of NP69‑LMP1Δ232‑351 cells was significantly decreased compared with cells expressing wild type LMP1 (LMP1WT; n=3; P<0.05); ii) 17 proteins exhibited differential protein expression (>2‑fold change) in NP69‑LMP1Δ232‑351 cells compared with NP69‑LMP1WT cells; and iii) LMP1WT was involved in activating the Janus kinase 3 (JAK3) promoter and regulating the expression of JAK3 protein, while LMP1Δ232‑351 was almost defective in ability to activate the JAK promoter. These results suggested that LMP1‑CTAR3 may be an important functional domain for regulating cell proliferation and protein expression in nasopharyngeal epithelial cells.

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1	Thompson MP and Kurzrock R: Epstein-Barr virus and cancer. Clin Cancer Res. 10:803–821. 2004. View Article : Google Scholar : PubMed/NCBI
2	Ma SD, Tsai MH, Romero-Masters JC, Ranheim EA, Huebner SM, Bristol JA, Delecluse HJ and Kenney SC: Latent membrane protein 1 (LMP1) and LMP2A collaborate to promote Epstein-Barr virus-induced B cell lymphomas in a cord blood-humanized mouse model but are not essential. J Virol. 91:e01928–16. 2017. View Article : Google Scholar : PubMed/NCBI
3	Pandya J and Walling DM: Oncogenic activity of Epstein-Barr virus latent membrane protein 1 (LMP-1) is down-regulated by lytic LMP-1. J Virol. 80:8038–8046. 2006. View Article : Google Scholar : PubMed/NCBI
4	Lavorgna A and Harhaj EW: EBV LMP1: New and shared pathways to NF-kB activation. Proc Natl Acad Sci USA. 109:2188–2189. 2012. View Article : Google Scholar : PubMed/NCBI
5	Ikeda O, Miyasaka Y, Yoshida R, Mizushima A, Oritani K, Sekine Y, Kuroda M, Yasui T, Fujimuro M, Muromoto R, et al: BS69 cooperates with TRAF3 in the regulation of Epstein-Barr virus-derived LMP1/CTAR1-induced NF-kappaB activation. FEBS Lett. 584:865–872. 2010. View Article : Google Scholar : PubMed/NCBI
6	Shkoda A, Town JA, Griese J, Romio M, Sarioglu H, Knöfel T, Giehler F and Kieser A: The germinal center kinase TNIK is required for canonical NF-κB and JNK signaling in B-cells by the EBV oncoprotein LMP1 and the CD40 receptor. PLoS Biol. 10:e10013762012. View Article : Google Scholar : PubMed/NCBI
7	Gires O, Kohlhuber F, Kilger E, Baumann M, Kieser A, Kaiser C, Zeidler R, Scheffer B, Ueffing M and Hammerschmidt W: Latent membrane protein 1 of Epstein-Barr virus interacts with JAK3 and activates STAT proteins. EMBO J. 18:3064–3073. 1999. View Article : Google Scholar : PubMed/NCBI
8	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
9	Farrell PJ: Epstein-Barr virus and cancer. Annu Rev Pathol. 14:29–53. 2019. View Article : Google Scholar : PubMed/NCBI
10	Kilger E, Kieser A, Baumann M and Hammerschmidt W: Epstein-Barr virus-mediated B-cell proliferation is dependent upon latent membrane protein 1, which simulates an activated CD40 receptor. EMBO J. 17:1700–1709. 1998. View Article : Google Scholar : PubMed/NCBI
11	Shair KH, Schnegg CI and Raab-Traub N: EBV latent membrane protein 1 effects on plakoglobin, cell growth, and migration. Cancer Res. 68:6997–7005. 2008. View Article : Google Scholar : PubMed/NCBI
12	Tu C, Zeng Z, Qi P, Li X, Guo C, Xiong F, Xiang B, Zhou M, Liao Q, Yu J, et al: Identification of genomic alterations in nasopharyngeal carcinoma and nasopharyngeal carcinoma-derived Epstein-Barr virus by whole-genome sequencing. Carcinogenesis. 39:1517–1528. 2018. View Article : Google Scholar : PubMed/NCBI
13	Tsao SW, Wang X, Liu Y, Cheung YC, Feng H, Zheng Z, Wong N, Yuen PW, Lo AK, Wong YC and Huang DP: Establishment of two immortalized nasopharyngeal epithelial cell lines using SV40 large T and HPV16E6/E7 viral oncogenes. Biochim Biophys Acta. 1590:150–158. 2002. View Article : Google Scholar : PubMed/NCBI
14	Lo AK, Liu Y, Wang XH, Huang DP, Yuen PW, Wong YC and Tsao GS: Alterations of biologic properties and gene expression in nasopharyngeal epithelial cells by the Epstein-Barr virus-encoded latent membrane protein 1. Lab Invest. 83:697–709. 2003. View Article : Google Scholar : PubMed/NCBI
15	Ahsan N, Kanda T, Nagashima K and Takada K: Epstein-Barr virus transforming protein LMP1 plays a critical role in virus production. J Virol. 79:4415–4424. 2005. View Article : Google Scholar : PubMed/NCBI
16	Xin B, He Z, Yang X, Chan CP, Ng MH and Cao L: TRADD domain of Epstein-Barr virus transforming protein LMP1 is essential for inducing immortalization and suppressing senescence of primary rodent fibroblasts. J Virol. 75:3010–3015. 2001. View Article : Google Scholar : PubMed/NCBI
17	Gallardo-Vera F, Diaz D, Tapia-Rodriguez M, Fortoul van der Goes T, Masso F, Rendon-Huerta E and Montaño LF: Vanadium pentoxide prevents NK-92MI cell proliferation and IFNg secretion through sustained JAK3 phosphorylation. J Immunotoxicol. 13:27–37. 2016. View Article : Google Scholar : PubMed/NCBI
18	Smirnova KV, Diduk SV, Senyuta NB and Gurtsevitch VE: Molecular biological properties of the Epstein-Barr virus LMP1 gene: Structure, function and polymorphism. Vopr Virusol. 60:5–13. 2015.(In Russian). PubMed/NCBI
19	Uchida J, Yasui T, Takaoka-Shichijo Y, Muraoka M, Kulwichit W, Raab-Traub N and Kikutani H: Mimicry of CD40 signals by Epstein-Barr virus LMP1 in B lymphocyte responses. Science. 286:300–303. 1999. View Article : Google Scholar : PubMed/NCBI
20	Zhou C, Dai X, Chen Y, Shen Y, Lei S, Xiao T, Bartfai T, Ding J and Wang MW: G protein-coupled receptor GPR160 is associated with apoptosis and cell cycle arrest of prostate cancer cells. Oncotarget. 7:12823–12839. 2016.PubMed/NCBI
21	He Y and Smith R: Nuclear functions of heterogeneous nuclear ribonucleoproteins A/B. Cell Mol Life Sci. 66:1239–1256. 2009. View Article : Google Scholar : PubMed/NCBI
22	He Y, Brown MA, Rothnagel JA, Saunders NA and Smith R: Roles of heterogeneous nuclear ribonucleoproteins A and B in cell proliferation. J Cell Sci. 118:3173–3183. 2005. View Article : Google Scholar : PubMed/NCBI
23	Lu HC, Ma J, Zhuang Z, Qiu F, Cheng HL and Shi JX: Exploring the regulatory role of isocitrate dehydrogenase mutant protein on glioma stem cell proliferation. Eur Rev Med Pharmacol Sci. 20:3378–3384. 2016.PubMed/NCBI
24	Bentz GL, Whitehurst CB and Pagano JS: Epstein-Barr virus latent membrane protein 1 (LMP1) C-terminal-activating region 3 contributes to LMP1-mediated cellular migration via its interaction with Ubc9. J Virol. 85:10144–10153. 2011. View Article : Google Scholar : PubMed/NCBI