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嗜铁素还原地杆菌对铬酸盐胁迫响应的分子动力学

Molecular dynamics of the Shewanella oneidensis response to chromate stress.

作者信息

Brown Steven D, Thompson Melissa R, Verberkmoes Nathan C, Chourey Karuna, Shah Manesh, Zhou Jizhong, Hettich Robert L, Thompson Dorothea K

机构信息

Environmental Sciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831, USA.

出版信息

Mol Cell Proteomics. 2006 Jun;5(6):1054-71. doi: 10.1074/mcp.M500394-MCP200. Epub 2006 Mar 8.

Abstract

Temporal genomic profiling and whole-cell proteomic analyses were performed to characterize the dynamic molecular response of the metal-reducing bacterium Shewanella oneidensis MR-1 to an acute chromate shock. The complex dynamics of cellular processes demand the integration of methodologies that describe biological systems at the levels of regulation, gene and protein expression, and metabolite production. Genomic microarray analysis of the transcriptome dynamics of midexponential phase cells subjected to 1 mm potassium chromate (K(2)CrO(4)) at exposure time intervals of 5, 30, 60, and 90 min revealed 910 genes that were differentially expressed at one or more time points. Strongly induced genes included those encoding components of a TonB1 iron transport system (tonB1-exbB1-exbD1), hemin ATP-binding cassette transporters (hmuTUV), TonB-dependent receptors as well as sulfate transporters (cysP, cysW-2, and cysA-2), and enzymes involved in assimilative sulfur metabolism (cysC, cysN, cysD, cysH, cysI, and cysJ). Transcript levels for genes with annotated functions in DNA repair (lexA, recX, recA, recN, dinP, and umuD), cellular detoxification (so1756, so3585, and so3586), and two-component signal transduction systems (so2426) were also significantly up-regulated (p < 0.05) in Cr(VI)-exposed cells relative to untreated cells. By contrast, genes with functions linked to energy metabolism, particularly electron transport (e.g. so0902-03-04, mtrA, omcA, and omcB), showed dramatic temporal alterations in expression with the majority exhibiting repression. Differential proteomics based on multidimensional HPLC-MS/MS was used to complement the transcriptome data, resulting in comparable induction and repression patterns for a subset of corresponding proteins. In total, expression of 2,370 proteins were confidently verified with 624 (26%) of these annotated as hypothetical or conserved hypothetical proteins. The initial response of S. oneidensis to chromate shock appears to require a combination of different regulatory networks that involve genes with annotated functions in oxidative stress protection, detoxification, protein stress protection, iron and sulfur acquisition, and SOS-controlled DNA repair mechanisms.

摘要

进行了时间基因组分析和全细胞蛋白质组分析,以表征金属还原菌——希瓦氏菌MR-1对急性铬酸盐冲击的动态分子反应。细胞过程的复杂动态需要整合在调控、基因和蛋白质表达以及代谢产物产生水平上描述生物系统的方法。对处于对数中期的细胞在暴露时间间隔为5、30、60和90分钟时施加1 mM铬酸钾(K₂CrO₄)后的转录组动态进行基因组微阵列分析,发现910个基因在一个或多个时间点有差异表达。强烈诱导的基因包括那些编码TonB1铁转运系统(tonB1-exbB1-exbD1)、血红素ATP结合盒转运蛋白(hmuTUV)、TonB依赖性受体以及硫酸盐转运蛋白(cysP、cysW-2和cysA-2)的基因,以及参与同化硫代谢的酶(cysC、cysN、cysD、cysH、cysI和cysJ)。在DNA修复(lexA、recX、recA、recN、dinP和umuD)、细胞解毒(so1756、so3585和so3586)以及双组分信号转导系统(so2426)中具有注释功能的基因转录水平,在暴露于Cr(VI)的细胞中相对于未处理细胞也显著上调(p < 0.05)。相比之下,与能量代谢相关的基因,特别是电子传递相关基因(如so0902-03-04、mtrA、omcA和omcB),其表达呈现出显著的时间变化,大多数表现为抑制。基于多维HPLC-MS/MS的差异蛋白质组学用于补充转录组数据,导致相应蛋白质子集中出现类似的诱导和抑制模式。总共可靠地验证了2370种蛋白质的表达,其中624种(26%)被注释为假设或保守假设蛋白质。希瓦氏菌对铬酸盐冲击的初始反应似乎需要不同调控网络相结合,这些网络涉及在氧化应激保护、解毒、蛋白质应激保护、铁和硫获取以及SOS控制的DNA修复机制中具有注释功能的基因。

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