Anomalies of the anaerobic tricarboxylic acid cycle in Shewanella oneidensis revealed by Tn-seq
Anomalies of the anaerobic tricarboxylic acid cycle in Shewanella oneidensis revealed by Tn-seq
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DOI：
10.1111/j.1365-2958.2012.08196.x
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发表时间：
2012-10-01
影响因子：
3.6
通讯作者：
Gralnick, Jeffrey A.
中科院分区：
文献类型：
--
作者：
Brutinel, Evan D.;Gralnick, Jeffrey A.
越来越便宜的测序与不断扩展的分子生物学工具箱相结合的可用性已经将经典的细菌遗传学带入了21世纪世纪。利用转座子诱变结合下一代高通量测序（Tn-seq）的全基因组遗传适合度分析有望彻底改变微生物代谢的系统水平分析。Tn-seq测量了经历纯化选择的异质突变库的实际成员的频率，以确定基因组中每个非必需基因对给定条件下生物体适应性的贡献。在这里，我们使用Tn-seq来评估革兰氏阴性？变形杆菌Shewanella oneidensis菌株MR-1。除了作为一个模式环境生物，有相当大的兴趣，使用S。oneidensis作为生物修复和生物技术的平台生物，需要对可能利用的代谢途径有一个完整的了解。我们的分析揭示了S.？oneidensis代谢被忽略了超过30年的经典遗传和系统水平的分析。我们报告的利用替代柠檬酸合酶和描述的S.？oneidensis厌氧三羧酸循环，在任何其他生物体中未报道，这可能是一种广泛的策略，微生物善于通过厌氧呼吸消散还原当量。
The availability of increasingly inexpensive sequencing combined with an ever-expanding molecular biology toolbox has transported classical bacterial genetics into the 21st century. Whole genome genetic fitness analysis using transposon mutagenesis combined with next-generation high-throughput sequencing (Tn-seq) promises to revolutionize systems level analysis of microbial metabolism. Tn-seq measures the frequency of actual members of a heterogeneous mutant pool undergoing purifying selection to determine the contribution of every non-essential gene in the genome to the fitness of an organism under a given condition. Here we use Tn-seq to assess gene function in the Gram negative ?-proteobacterium Shewanella oneidensis strain MR-1. In addition to being a model environmental organism, there is considerable interest in using S.?oneidensis as a platform organism for bioremediation and biotechnology, necessitating a complete understanding of the metabolic pathways that may be utilized. Our analysis reveals unique aspects of S.?oneidensis metabolism overlooked by over 30 years of classical genetic and systems level analysis. We report the utilization of an alternative citrate synthase and describe a dynamic branching of the S.?oneidensis anaerobic tricarboxylic acid cycle, unreported in any other organism, which may be a widespread strategy for microbes adept at dissipating reducing equivalents via anaerobic respiration.