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首页> 外文期刊>Applied Microbiology >Determining Roles of Accessory Genes in Denitrification by Mutant Fitness Analyses
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Determining Roles of Accessory Genes in Denitrification by Mutant Fitness Analyses

机译：通过突变适应性分析确定辅助基因在反硝化中的作用

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Enzymes of the denitrification pathway play an important role in the global nitrogen cycle, including release of nitrous oxide, an ozone-depleting greenhouse gas. In addition, nitric oxide reductase, maturation factors, and proteins associated with nitric oxide detoxification are used by pathogens to combat nitric oxide release by host immune systems. While the core reductases that catalyze the conversion of nitrate to dinitrogen are well understood at a mechanistic level, there are many peripheral proteins required for denitrification whose basic function is unclear. A bar-coded transposon DNA library from Pseudomonas stutzeri strain RCH2 was grown under denitrifying conditions, using nitrate or nitrite as an electron acceptor, and also under molybdenum limitation conditions, with nitrate as the electron acceptor. Analysis of sequencing results from these growths yielded gene fitness data for 3,307 of the 4,265 protein-encoding genes present in strain RCH2. The insights presented here contribute to our understanding of how peripheral proteins contribute to a fully functioning denitrification pathway. We propose a new low-affinity molybdate transporter, OatABC, and show that differential regulation is observed for two MoaA homologs involved in molybdenum cofactor biosynthesis. We also propose that NnrS may function as a membrane-bound NO sensor. The dominant HemN paralog involved in heme biosynthesis is identified, and a CheR homolog is proposed to function in nitrate chemotaxis. In addition, new insights are provided into nitrite reductase redundancy, nitric oxide reductase maturation, nitrous oxide reductase maturation, and regulation.
机译：反硝化途径的酶在全球氮循环中起重要作用，包括释放一氧化二氮（一种消耗臭氧的温室气体）。另外，病原体使用一氧化氮还原酶，成熟因子和与一氧化氮解毒相关的蛋白质来抵抗宿主免疫系统释放的一氧化氮。虽然从机理上可以很好地理解催化硝酸盐转化为二氮的核心还原酶，但反硝化所需的许多外围蛋白尚不清楚，其基本功能尚不清楚。在反硝化条件下，使用硝酸盐或亚硝酸盐作为电子受体，并且在钼限制条件下，以硝酸盐作为电子受体，生长来自斯氏假单胞菌RCH2的条形码转座子DNA文库。对这些生长的测序结果进行的分析得出了菌株RCH2中存在的4,265个蛋白质编码基因中的3,307个的基因适合度数据。此处介绍的见解有助于我们了解外围蛋白如何促进功能全面的反硝化途径。我们提出了一种新的低亲和力钼酸盐转运蛋白OatABC，并表明对参与钼辅助因子生物合成的两个MoaA同源物观察到差异调节。我们还建议NnrS可以充当膜结合的NO传感器。确定了占主导地位的血红素生物合成涉及的HemN旁系同源物，并提出了CheR同源物在硝酸盐趋化作用中发挥作用。此外，还提供了有关亚硝酸还原酶冗余，一氧化氮还原酶成熟，一氧化二氮还原酶成熟和调节的新见解。

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