Department of Microbiology and Immunology, Otago School of Medical Sciences, University of Otago, Dunedin, New Zealand.
BMC Microbiol. 2009 Dec 10;9:254. doi: 10.1186/1471-2180-9-254.
Mycobacteria have been shown to contain an apparent redundancy of high-affinity phosphate uptake systems, with two to four copies of such systems encoded in all mycobacterial genomes sequenced to date. In addition, all mycobacteria also contain at least one gene encoding the low-affinity phosphate transporter, Pit. No information is available on a Pit system from a Gram-positive microorganism, and the importance of this system in a background of multiple other phosphate transporters is unclear.
The aim of this study was to determine the physiological role of the PitA phosphate transporter in Mycobacterium smegmatis. Expression of pitA was found to be constitutive under a variety of growth conditions. An unmarked deletion mutant in pitA of M. smegmatis was created. The deletion did not affect in vitro growth or phosphate uptake of M. smegmatis. Expression of the high-affinity transporters, PstSCAB and PhnDCE, was increased in the pitA deletion strain.
PitA is the only low-affinity phosphate transport system annotated in the genome of M. smegmatis. The lack of phenotype of the pitA deletion strain shows that this system is dispensable for in vitro growth of this organism. However, increased expression of the remaining phosphate transporters in the mutant indicates a compensatory mechanism and implies that PitA is indeed used for the uptake of phosphate in M. smegmatis.
已证实分枝杆菌含有高亲和力磷酸盐摄取系统的明显冗余,迄今为止测序的所有分枝杆菌基因组中都有两个到四个此类系统的副本。此外,所有分枝杆菌还至少含有一个编码低亲和力磷酸盐转运体 Pit 的基因。尚未获得革兰氏阳性微生物的 Pit 系统的信息,并且在存在多个其他磷酸盐转运体的背景下该系统的重要性尚不清楚。
本研究的目的是确定 Mycobacterium smegmatis 中 PitA 磷酸盐转运体的生理作用。发现 pitA 的表达在各种生长条件下都是组成型的。创建了 M. smegmatis 中 pitA 的无标记缺失突变体。该缺失突变不影响 M. smegmatis 的体外生长或磷酸盐摄取。高亲和力转运体 PstSCAB 和 PhnDCE 的表达在 pitA 缺失菌株中增加。
PitA 是 M. smegmatis 基因组中注释的唯一低亲和力磷酸盐转运系统。pitA 缺失菌株的表型缺失表明该系统对于该生物体的体外生长并非必需。然而,突变体中剩余磷酸盐转运体的表达增加表明存在补偿机制,这意味着 PitA 确实用于 M. smegmatis 中磷酸盐的摄取。