Effects of HIV-1 Vpr protein on cell viability and cell cycle of mouse neuroblastoma N2a cells
10.3760/cma.j.issn.1003-9279.2019.05.010
- VernacularTitle: HIV-1 Vpr蛋白对小鼠神经瘤母细胞N2a活性及细胞周期的影响
- Author:
Xinyue CAO
1
;
Xiaoyu SHAN
1
;
Wenhui ZHENG
1
;
Shuaizhi GUO
1
;
Zeming QIN
2
;
Hongling WEN
1
;
Li ZHAO
1
Author Information
1. Department of Hygienic Microbiology, School of Public Health, Shandong University, Jinan 250012, China
2. Experimental Teaching Center of Preventive Medicine, School of Public Health, Shandong University, Jinan 250012, China
- Publication Type:Journal Article
- Keywords:
Human immunodeficiency virus type 1;
Vpr protein;
HIV associated dementia;
cell G2 arrest
- From:
Chinese Journal of Experimental and Clinical Virology
2019;33(5):489-494
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the toxic effect of HIV-1 Vpr protein on neurons.
Methods:HIV-1 vpr gene was amplified by nested PCR in four parts of peripheral spleen (SPL) and central nervous tissue meninges (MG) of HIV-associated dementia (HAD) patients and non-HAD patients. Eukaryotic expression vector pEGFP-N1-vpr was constructed. The gene sequence and key amino acid sites were analyzed by BLAST and MEGA6. The expression of Vpr protein in N2a cells was detected by Western-blotting. The effects of Vpr proteins from different sources on the activity and cell cycle of N2a cells were studied by flow cytometry.
Results:HIV-1 vpr gene was successfully amplified by PCR. Sequence analysis showed that the vpr gene sequence belonged to HIV-1B subtype. There were amino acid mutations at C-terminal 84, 86 and 87 sites of central Vpr protein from HAD and non-HAD patients. Vpr protein could inhibit the activity of nerve cells, leading to G2 phase arrest. Different sources of Vpr had different intensity of action. Compared with other groups, Vpr protein from the meninges of HAD patients showed stronger inhibition of cell activity and G2 phase arrest ability.
Conclusions:Variations in key amino acid sites of Vpr protein could cause significant changes in its biological functions, and its significance in the pathogenesis of HAD remains to be further studied.
