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首页> 外文期刊>Microbiology >Deletion of SenX3–RegX3, a key two-component regulatory system of Mycobacterium smegmatis, results in growth defects under phosphate-limiting conditions
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Deletion of SenX3–RegX3, a key two-component regulatory system of Mycobacterium smegmatis, results in growth defects under phosphate-limiting conditions

机译：删除Senx3-Regx3，鉴于磷酸盐限制条件下的生长缺陷的关键双组分调节系统

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摘要

Two component regulatory systems are key elements in the control of bacterial gene expression in response to environmental perturbations. The SenX3–RegX3 system is implicated in the control of phosphate uptake in Mycobacterium smegmatis and Mycobacterium tuberculosis. regX3 is reported to be essential in M. smegmatis, but not in M. tuberculosis. We attempted to construct complete senX3–regX3 operon deletion strains of M. smegmatis; initially we found that the operon could only be deleted when another functional copy was provided. Using a strain in which the only functional copy of the operon was present on an integrating plasmid, we attempted to replace the functional copy with an empty vector. Surprisingly, we obtained strains in which the functional copy had been deleted from the chromosome at a low frequency. We deleted the senX3 gene in a similar fashion, but it was not possible to delete regX3 alone. To identify possible compensatory mutations we sequenced the whole genome of two deletion strains and the wild-type. A synonymous single nucleotide polymorphism (SNP) in a lipoprotein was found in all deletion strains, but not the parental strains, and a frameshift mutation in nhaA was identified in three of the four deletion strains. Operon deletion strains were more sensitive to phosphate limitation, showing a reduced ability to grow at lower phosphate concentrations. The M. tuberculosis operon was able to functionally complement the growth phenotype in M. smegmatis under phosphate-replete conditions, but not under low phosphate conditions, reinforcing the difference between the two species. Our data show that, in contrast with previous reports, it is possible to delete the operon in M. smegmatis, possibly due to the accumulation of compensatory mutations, and that the deletion does affect growth in phosphate.
机译：两个组分调节系统是控制细菌基因表达的关键元素，以应对环境扰动。 Senx3-Regx3系统涉及控制分枝杆菌和结核分枝杆菌中的磷酸盐摄取。据报道，Regx3在M. Smogmatis中是必需的，但在肺结核中没有。我们试图构建完整的Senx3-Regx3操纵子删除M. Smogmatis;最初我们发现，只有在提供了另一个功能副本时才能删除操纵子。使用其中唯一在整合质粒上存在操纵子的功能拷贝的菌株，我们试图用空向量取代功能副本。令人惊讶的是，我们获得了在低频率下从染色体中删除功能拷贝的菌株。我们以类似的方式删除了SENX3基因，但无法单独删除REGX3。为了鉴定可能的补偿性突变，我们测序了两种缺失菌株的全基因组和野生型。在所有缺失菌株中发现脂蛋白中的同义单核苷酸多态性（SNP），但不是父母菌株，并且在四种缺失菌株中的三个中鉴定了NHAA中的帧突变突变。操纵子缺失菌株对磷酸盐限制更敏感，显示出在较低的磷酸盐浓度下生长的能力。在磷酸盐 - 新鲜条件下，M.结核术术能够在磷酸盐条件下在M. Smogmatis中的生长表型补充，但在低磷酸盐条件下，加强了两种物种之间的差异。我们的数据表明，与先前的报告相比，可以删除M. Smogmatis的操纵子，可能是由于补偿性突变的积累，并且缺失会影响磷酸盐的生长。

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