HIV-1 Vpr Modulates Macrophage Metabolic Pathways: A SILAC-Based Quantitative Analysis

被引:78
作者
Barrero, Carlos A. [1 ,3 ]
Datta, Prasun K. [2 ,3 ]
Sen, Satarupa [2 ,4 ]
Deshmane, Satish [2 ,3 ]
Amini, Shohreh [2 ,3 ,4 ]
Khalili, Kamel [2 ,3 ]
Merali, Salim [1 ,3 ]
机构
[1] Temple Univ, Sch Med, Fels Inst, Dept Biochem, Philadelphia, PA 19122 USA
[2] Temple Univ, Sch Med, Dept Neurosci, Philadelphia, PA 19122 USA
[3] Temple Univ, Sch Med, Ctr Neurovirol, Philadelphia, PA 19122 USA
[4] Temple Univ, Dept Biol, Philadelphia, PA 19122 USA
来源
PLOS ONE | 2013年 / 8卷 / 07期
基金
美国国家卫生研究院;
关键词
IMMUNODEFICIENCY-VIRUS TYPE-1; OXYGEN-REGULATED PROTEIN; PYRUVATE-KINASE M2; GENE-TRANSCRIPTION; PROTEOMIC ANALYSIS; T-CELLS; REPLICATION; EXPRESSION; INFECTION; APOPTOSIS;
D O I
10.1371/journal.pone.0068376
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Human immunodeficiency virus type 1 encoded viral protein Vpr is essential for infection of macrophages by HIV-1. Furthermore, these macrophages are resistant to cell death and are viral reservoir. However, the impact of Vpr on the macrophage proteome is yet to be comprehended. The goal of the present study was to use a stable-isotope labeling by amino acids in cell culture (SILAC) coupled with mass spectrometry-based proteomics approach to characterize the Vpr response in macrophages. Cultured human monocytic cells, U937, were differentiated into macrophages and transduced with adenovirus construct harboring the Vpr gene. More than 600 proteins were quantified in SILAC coupled with LC-MS/MS approach, among which 136 were significantly altered upon Vpr overexpression in macrophages. Quantified proteins were selected and clustered by biological functions, pathway and network analysis using Ingenuity computational pathway analysis. The proteomic data illustrating increase in abundance of enzymes in the glycolytic pathway (pentose phosphate and pyruvate metabolism) was further validated by western blot analysis. In addition, the proteomic data demonstrate down regulation of some key mitochondrial enzymes such as glutamate dehydrogenase 2 (GLUD2), adenylate kinase 2 (AK2) and transketolase (TKT). Based on these observations we postulate that HIV-1 hijacks the macrophage glucose metabolism pathway via the Vpr-hypoxia inducible factor 1 alpha (HIF-1 alpha) axis to induce expression of hexokinase (HK), glucose-6-phosphate dehyrogenase (G6PD) and pyruvate kinase muscle type 2 (PKM2) that facilitates viral replication and biogenesis, and long-term survival of macrophages. Furthermore, dysregulation of mitochondrial glutamate metabolism in macrophages can contribute to neurodegeneration via neuroexcitotoxic mechanisms in the context of NeuroAIDS.
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页数:11
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