Disruption of the pdhB pyruvate dehydrogenase [corrected] gene affects colony morphology, in vitro growth and cell invasiveness of Mycoplasma agalactiae.
Disruption of the pdhB pyruvate dehydrogenase [corrected] gene affects colony morphology, in vitro growth and cell invasiveness of Mycoplasma agalactiae.
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DOI：
10.1371/journal.pone.0119706
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Chopra-Dewasthaly R
中科院分区：
文献类型：
--
作者：
Hegde S;Rosengarten R;Chopra-Dewasthaly R
细菌对有效底物的利用、代谢潜力和生长速率在其致病性中起重要作用。本研究集中于无乳支原体，它通过其对小反刍动物传染性无乳的贡献，通过未知的机制，造成重大的经济损失。这种知识的缺乏主要是由于其苛刻的生长要求和可用于其操作和分析的遗传工具的稀缺。转座子诱变M.无乳型菌株PG 2导致整个基因组的几个破坏。发现在体外生长缺陷的突变体在pdhB基因中具有转座子插入，该基因编码丙酮酸脱氢酶复合物的组分。这种生长差异在活跃分裂的对数期非常显著，但随着细胞接近稳定期，观察到逐渐恢复。该突变体还表现出与野生型菌株PG 2相比不同且更小的菌落形态。为了互补，将pdhAB克隆到新构建的互补载体中的强vpma启动子下游和lacZ报告基因上游。当用该载体转化时，pdhB突变体恢复其正常生长和菌落形态。有趣的是，pdhB突变体在HeLa细胞中的侵袭性也显著降低，如双重免疫荧光染色所示。这种缺陷在补充菌株中得到恢复，其侵袭性与PG 2相当。综上所述，这些数据表明，丙酮酸脱氢酶可能是一个重要的球员在感染和定植的M。无乳。
The utilization of available substrates, the metabolic potential and the growth rates of bacteria can play significant roles in their pathogenicity. This study concentrates on Mycoplasma agalactiae, which causes significant economic losses through its contribution to contagious agalactia in small ruminants by as yet unknown mechanisms. This lack of knowledge is primarily due to its fastidious growth requirements and the scarcity of genetic tools available for its manipulation and analysis. Transposon mutagenesis of M. agalactiae type strain PG2 resulted in several disruptions throughout the genome. A mutant defective in growth in vitro was found to have a transposon insertion in the pdhB gene, which encodes a component of the pyruvate dehydrogenase complex. This growth difference was quite significant during the actively dividing logarithmic phase but a gradual recovery was observed as the cells approached stationary phase. The mutant also exhibited a different and smaller colony morphology compared to the wild type strain PG2. For complementation, pdhAB was cloned downstream of a strong vpma promoter and upstream of a lacZ reporter gene in a newly constructed complementation vector. When transformed with this vector the pdhB mutant recovered its normal growth and colony morphology. Interestingly, the pdhB mutant also had significantly reduced invasiveness in HeLa cells, as revealed by double immunofluorescence staining. This deficiency was recovered in the complemented strain, which had invasiveness comparable to that of PG2. Taken together, these data indicate that pyruvate dehydrogenase might be an important player in infection with and colonization by M. agalactiae.
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影响因子：
3.1
作者：
Cieri, MV;Mayer-Hamblett, N;Burns, JL
通讯作者：
Burns, JL
影响因子：
3.2
作者：
Dybvig, Kevin;Cao, Z.;Yu, Huilan
通讯作者：
Yu, Huilan
影响因子：
4.4
作者：
Maglennon GA;Cook BS;Matthews D;Deeney AS;Bossé JT;Langford PR;Maskell DJ;Tucker AW;Wren BW;Rycroft AN;BRaDP1T consortium
通讯作者：
BRaDP1T consortium
影响因子：
2.8
作者：
Lee, S. -W;Browning, G. F.;Markham, P. F.
通讯作者：
Markham, P. F.
影响因子：
2.2
作者：
Li, Jiahe;Zhang, Jixiang;Li, Jinxiang
通讯作者：
Li, Jinxiang