Quantitative Proteomics Reveals Novel Insights into Isoniazid Susceptibility in Mycobacteria Mediated by a Universal Stress Protein
Quantitative Proteomics Reveals Novel Insights into Isoniazid Susceptibility in Mycobacteria Mediated by a Universal Stress Protein
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定量蛋白质组学揭示了通用应激蛋白介导的分枝杆菌异烟肼敏感性的新见解
DOI：
10.1021/pr5011058
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发表时间：
2015-03-01
影响因子：
4.4
通讯作者：
Mi, Kaixia
中科院分区：
文献类型：
--
作者：
Hu, Xinling;Li, Xiaojing;Mi, Kaixia
结核病（TB）是由古老的病原体结核分枝杆菌引起的，是世界上最严重的传染病之一。异烟肼（INH）是治疗活动性和潜伏性结核病的重要一线药物。 INH 耐药性是结核病治疗中日益严重的问题。然而，人们对 INH 的表型耐药性知之甚少。在本研究中，我们构建了一株过度表达潜伏期相关通用应激蛋白（USP）的牛分枝杆菌BCG菌株BCG_2013，并将该菌株命名为BCG-2013。与野生型菌株 BCG-pMV261 相比，BCG_2013 过表达增加了对 INH 的敏感性。定量蛋白质组分析显示，在已鉴定的 1500 个蛋白质中，BCG_2013 过表达导致 50 个蛋白质上调，26 个蛋白质下调。通过 qPCR 观察并证实了 BCG-2013 中过氧化氢酶-过氧化物酶 KatG 表达的上调，而其他 INH 耐药相关蛋白的表达没有变化。此外，还观察到分枝杆菌持久性调节因子 MprA 及其调节蛋白的差异表达。在 Wayne 休眠模型中，BCG_2013 和 katG mRNA 水平增加，而 MprA mRNA 水平下降。综上所述，我们的结果表明，BCG_2013 水平升高诱导 KatG 水平升高是分枝杆菌对 INH 表型敏感性的基础。
Tuberculosis (TB) is caused by the ancient pathogen, Mycobacterium tuberculosis, and is one of the most serious infectious diseases in the world. Isoniazid (INH) is an important first-line drug for the treatment of active and latent TB. INH resistance is an increasing problem in the treatment of TB. Phenotypic resistance to INH, however, is poorly understood. In this study, we constructed a strain of Mycobacterium bovis BCG that overexpresses the latency-related universal stress protein (USP), BCG_2013, and designated this strain BCG-2013. BCG_2013 overexpression increased susceptibility to INH compared with that of the wild-type strain, BCG-pMV261. Quantitative proteomic analysis revealed that BCG_2013 overexpression resulted in the upregulation of 50 proteins and the downregulation of 26 proteins among the 1500 proteins identified. Upregulation of catalase-peroxidase KatG expression in BCG-2013 was observed and confirmed by qPCR, whereas expression of other INH resistance-related proteins did not change. In addition, differential expression of the mycobacterial persistence regulator MprA and its regulatory proteins was observed. BCG_2013 and katG mRNA levels increased in a Wayne dormancy model, whereas MprA mRNA levels decreased. Taken together, our results suggest that the increase in KatG levels induced by increased BCG_2013 levels underlies the phenotypic susceptibility of mycobacteria to INH.