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利用 DNA 微阵列技术研究氧限制、缺乏细胞色素 bc(1)复合物和低 pH 对氧化葡萄糖酸杆菌 621H 全局基因表达的影响。

Influence of oxygen limitation, absence of the cytochrome bc(1) complex and low pH on global gene expression in Gluconobacter oxydans 621H using DNA microarray technology.

机构信息

Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich, D-52425 Jülich, Germany.

出版信息

J Biotechnol. 2012 Feb 10;157(3):359-72. doi: 10.1016/j.jbiotec.2011.12.020. Epub 2011 Dec 29.

Abstract

The genome-wide transcriptional responses of the strictly aerobic α-proteobacterium Gluconobacter oxydans 621H to oxygen limitation, to the absence of the cytochrome bc(1) complex, and to low pH were studied using DNA microarray analyses. Oxygen limitation caused expression changes of 486 genes, representing 20% of the chromosomal genes. Genes with an increased mRNA level included those for terminal oxidases, the cytochrome bc(1) complex, transhydrogenase, two alcohol dehydrogenases, heme biosynthesis, PTS proteins, proteins involved in cyclic diGMP synthesis and degradation, two sigma factors, flagella and chemotaxis proteins, several stress proteins, and a putative exporter protein. The downregulated genes comprised those for respiratory dehydrogenases, enzymes of central metabolism, PQQ biosynthesis, outer membrane receptors, Sec proteins, and proteins involved in transcription and translation. A ΔqrcABC mutant of G. oxydans showed a growth defect during cultivation on mannitol at pH 4 under oxygen saturation. Comparison of the transcriptomes of this mutant versus the wild type under these conditions revealed 51 differentially expressed genes. Interestingly, almost all of the 45 genes with increased expression in the ΔqrcABC mutant at pH 4 were also upregulated in the wild type grown at pH 6 under oxygen limitation. These results support an active role of the cytochrome bc(1) complex in G. oxydans respiration. The transcriptome comparison of G. oxydans wild type at pH 4 versus pH 6 in mannitol medium under oxygen-saturated conditions uncovered only 72 differentially expressed genes. The 35 upregulated genes included those for cytochrome bd oxidase, major polyol dehydrogenase, iron storage and oxidative stress proteins. Among the 37 downregulated genes were some encoding enzymes dealing with carbon dioxide, such as biotin carboxylase, biotin carboxyl carrier protein, and carboanhydrase. These results give first insights into global transcriptional responses of G. oxydans.

摘要

采用 DNA 微阵列分析研究了严格需氧的α变形菌氧化葡萄糖酸杆菌 621H 对氧限制、缺乏细胞色素 bc(1) 复合物和低 pH 的全基因组转录反应。氧限制导致 486 个基因的表达变化,占染色体基因的 20%。mRNA 水平增加的基因包括末端氧化酶、细胞色素 bc(1) 复合物、转氢酶、两种醇脱氢酶、血红素生物合成、PTS 蛋白、参与环二鸟苷酸合成和降解的蛋白、两个σ因子、鞭毛和趋化蛋白、几种应激蛋白和一个假定的外排蛋白。下调的基因包括呼吸脱氢酶、中心代谢酶、PQQ 生物合成、外膜受体、Sec 蛋白和涉及转录和翻译的蛋白。在 pH 4 下用甘露醇培养时,G. oxydans 的ΔqrcABC 突变体表现出生长缺陷,在氧气饱和下。与野生型相比,在这些条件下,突变体和野生型的转录组比较显示 51 个差异表达基因。有趣的是,在 pH 4 下ΔqrcABC 突变体中表达增加的几乎所有 45 个基因在野生型中也在 pH 6 下氧气限制下被上调。这些结果支持细胞色素 bc(1) 复合物在 G. oxydans 呼吸中的积极作用。在氧气饱和条件下,G. oxydans 野生型在 pH 4 与 pH 6 的甘露醇培养基中的转录组比较仅发现 72 个差异表达基因。上调的 35 个基因包括细胞色素 bd 氧化酶、主要多元醇脱氢酶、铁储存和氧化应激蛋白。下调的 37 个基因中有些编码与二氧化碳有关的酶,如生物素羧化酶、生物素羧基载体蛋白和碳酸酐酶。这些结果首次深入了解了 G. oxydans 的全局转录反应。

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