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氧化应激作用于特殊的膜蛋白,降低了番茄丁香假单胞菌 pv 的生存能力。

Oxidative stress acts on special membrane proteins to reduce the viability of Pseudomonas syringae pv tomato.

机构信息

Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.

出版信息

J Proteome Res. 2012 Oct 5;11(10):4927-38. doi: 10.1021/pr300446g. Epub 2012 Sep 10.

Abstract

Reactive oxygen species (ROS) play a vital role in reducing the viability of invading pathogens during plant-pathogen interactions. To understand how oxidative stress caused by ROS reduces cell viability, it is important to identify the proteins affected by ROS. In the present study, we investigated the changes in the expression of proteins from the outer and inner membrane fractions in Pseudomonas syringae pv tomato DC3000 under oxidative stress through membrane subproteomics. A total number of 17 differentially expressed proteins from the outer and inner membrane fractions were identified, among which 11 proteins belong to transporters, such as porins and ABC transporters. Their abundance was all decreased under oxidative stress, indicating that transporters are likely to be affected by oxidative stress. The function of two identified transporters was further characterized by constructing their gene mutant and overexpression strains. We found that mutation of one transporter gene PSPTO_1720 rendered Pseudomonas more sensitive to oxidative stress, whereas overexpression of this gene made the strain more resistant. By comparison, the mutant and overexpression strains of another transporter gene PSPTO_2152 exhibited the same sensitivity to oxidative stress compared with the wild-type. Our data suggest that oxidative stress reduces the viability of bacterial cells by acting on special transporters.

摘要

活性氧(ROS)在植物-病原体相互作用过程中降低入侵病原体的生存能力方面起着至关重要的作用。为了了解 ROS 引起的氧化应激如何降低细胞活力,识别受 ROS 影响的蛋白质是很重要的。在本研究中,我们通过膜亚蛋白质组学研究了在番茄溃疡病菌 DC3000 中 ROS 对其外膜和内膜蛋白的影响。在外膜和内膜蛋白中鉴定出了 17 种差异表达蛋白,其中 11 种蛋白属于转运蛋白,如孔蛋白和 ABC 转运蛋白。在氧化应激下,它们的丰度都降低了,这表明转运蛋白可能受到氧化应激的影响。通过构建其基因突变体和过表达菌株,进一步对两种鉴定出的转运蛋白的功能进行了表征。我们发现,一个转运蛋白基因 PSPTO_1720 的突变使假单胞菌对氧化应激更敏感,而该基因的过表达则使菌株更具抗性。相比之下,另一个转运蛋白基因 PSPTO_2152 的突变体和过表达菌株与野生型相比,对氧化应激的敏感性相同。我们的数据表明,氧化应激通过作用于特殊的转运蛋白来降低细菌细胞的活力。

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