Isolation and characterization of a suppressor mutation that restores Myxococcus xanthus exopolysaccharide production.

Isolation and characterization of a suppressor mutation that restores Myxococcus xanthus exopolysaccharide production.
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DOI:
10.1099/mic.0.031070-0
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发表时间:
2009-11
期刊:
Microbiology (Reading, England)
影响因子:
--
通讯作者:
Yang Z
Yang Z
中科院分区:
其他
文献类型:
--
作者:
Black WP;Xu Q;Cadieux CL;Suh SJ;Shi W;Yang Z

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黄粘球菌是一种革兰氏阴性土壤细菌,当营养变得有限时,它会经历多细胞发育。聚集是发育过程的一部分,需要这种生物的表面运动性。黄豆运动的一个组成部分,社会(S)滑翔运动,使细胞在近距离的物理接近运动。先前的研究表明,细胞表面相关的外多糖(EPS)对S的运动至关重要,并且Dif蛋白形成了一种趋化样途径,调节黄原草中EPS的产生。DifA是Dif系统中甲基接受趋化蛋白(MCPs)的同源物,是EPS产生、S运动和发育所必需的。在这项研究中,分离了difA缺失的自发外基因抑制因子,以确定EPS产生的其他调节因子。抑制基因突变是在cheW7趋化基因簇上插入一个碱基对。进一步的研究表明,cheW7的突变可能导致Mcp7与DifC (CheW-like)和DifE (CheA-like)相互作用,以重建在缺乏DifA的情况下调节EPS产生的功能途径。此外,在difA+背景下,发现cheW7突变可以部分抑制EPS产生中的pilA突变。从pilA cheW7双突变体中进一步删除difA导致产生野生型水平EPS的三突变体,这表明difA (mcp样)和Mcp7在调节EPS产生时竞争与DifC和DifE的相互作用。
Myxococcus xanthus, a Gram-negative soil bacterium, undergoes multicellular development when nutrients become limiting. Aggregation, which is part of the developmental process, requires the surface motility of this organism. One component of M. xanthus motility, the social (S) gliding motility, enables the movement of cells in close physical proximity. Previous studies demonstrated that the cell-surface associated exopolysaccharide (EPS) is essential for S motility and the Dif proteins form a chemotaxis-like pathway that regulates EPS production in M. xanthus. DifA, a homologue of methyl-accepting chemotaxis proteins (MCPs) in the Dif system, is required for EPS production, S motility and development. In this study, a spontaneous extragenic suppressor of a difA deletion was isolated in order to identify additional regulators of EPS production. The suppressor mutation was found to be a single base-pair insertion in cheW7 at the che7 chemotaxis gene cluster. Further examination indicated that mutations in cheW7 may lead to the interaction of Mcp7 with DifC (CheW-like) and DifE (CheA-like) to reconstruct a functional pathway to regulate EPS production in the absence of DifA. In addition, the cheW7 mutation was found to partially suppress a pilA mutation in EPS production in a difA+ background. Further deletion of difA from the pilA cheW7 double mutant resulted in a triple mutant that produced wild-type levels of EPS, implying that DifA (MCP-like) and Mcp7 compete for interactions with DifC and DifE in the modulation of EPS production.
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