Clark M E, He Q, He Z, Huang K H, Alm E J, Wan X-F, Hazen T C, Arkin A P, Wall J D, Zhou J-Z, Fields M W
Department of Microbiology, Miami University, Oxford, OH 45056, USA.
Appl Environ Microbiol. 2006 Aug;72(8):5578-88. doi: 10.1128/AEM.00284-06.
Desulfovibrio vulgaris was cultivated in a defined medium, and biomass was sampled for approximately 70 h to characterize the shifts in gene expression as cells transitioned from the exponential to the stationary phase during electron donor depletion. In addition to temporal transcriptomics, total protein, carbohydrate, lactate, acetate, and sulfate levels were measured. The microarray data were examined for statistically significant expression changes, hierarchical cluster analysis, and promoter element prediction and were validated by quantitative PCR. As the cells transitioned from the exponential phase to the stationary phase, a majority of the down-expressed genes were involved in translation and transcription, and this trend continued at the remaining times. There were general increases in relative expression for intracellular trafficking and secretion, ion transport, and coenzyme metabolism as the cells entered the stationary phase. As expected, the DNA replication machinery was down-expressed, and the expression of genes involved in DNA repair increased during the stationary phase. Genes involved in amino acid acquisition, carbohydrate metabolism, energy production, and cell envelope biogenesis did not exhibit uniform transcriptional responses. Interestingly, most phage-related genes were up-expressed at the onset of the stationary phase. This result suggested that nutrient depletion may affect community dynamics and DNA transfer mechanisms of sulfate-reducing bacteria via the phage cycle. The putative feoAB system (in addition to other presumptive iron metabolism genes) was significantly up-expressed, and this suggested the possible importance of Fe2+ acquisition under metal-reducing conditions. The expression of a large subset of carbohydrate-related genes was altered, and the total cellular carbohydrate levels declined during the growth phase transition. Interestingly, the D. vulgaris genome does not contain a putative rpoS gene, a common attribute of the delta-Proteobacteria genomes sequenced to date, and the transcription profiles of other putative rpo genes were not significantly altered. Our results indicated that in addition to expected changes (e.g., energy conversion, protein turnover, translation, transcription, and DNA replication and repair), genes related to phage, stress response, carbohydrate flux, the outer envelope, and iron homeostasis played important roles as D. vulgaris cells experienced electron donor depletion.
将普通脱硫弧菌在限定培养基中培养,在大约70小时内对生物量进行采样,以表征细胞在电子供体耗尽期间从指数生长期过渡到稳定期时基因表达的变化。除了时间转录组学外,还测量了总蛋白、碳水化合物、乳酸、乙酸盐和硫酸盐水平。对微阵列数据进行统计显著表达变化检查、层次聚类分析和启动子元件预测,并通过定量PCR进行验证。随着细胞从指数生长期过渡到稳定期,大多数下调表达的基因参与翻译和转录,并且这种趋势在剩余时间持续。随着细胞进入稳定期,细胞内运输和分泌、离子运输以及辅酶代谢的相对表达普遍增加。正如预期的那样,DNA复制机制下调表达,而参与DNA修复的基因在稳定期表达增加。参与氨基酸获取、碳水化合物代谢、能量产生和细胞包膜生物合成的基因没有表现出一致的转录反应。有趣的是,大多数与噬菌体相关的基因在稳定期开始时上调表达。这一结果表明,营养物质耗尽可能通过噬菌体循环影响硫酸盐还原细菌的群落动态和DNA转移机制。假定的feoAB系统(以及其他推测的铁代谢基因)显著上调表达,这表明在金属还原条件下获取Fe2+可能具有重要意义。一大类与碳水化合物相关的基因表达发生改变,并且在生长阶段转变期间细胞总碳水化合物水平下降。有趣的是,普通脱硫弧菌基因组不包含假定的rpoS基因,这是迄今为止已测序的δ-变形菌基因组的一个共同特征,并且其他假定的rpo基因的转录谱没有显著改变。我们的结果表明,除了预期的变化(如能量转换、蛋白质周转、翻译、转录以及DNA复制和修复)外,与噬菌体、应激反应、碳水化合物通量、外膜和铁稳态相关的基因在普通脱硫弧菌细胞经历电子供体耗尽时发挥了重要作用。