Regulon controlled by the GppX hybrid two component system in Porphyromonas gingivalis.
Regulon controlled by the GppX hybrid two component system in Porphyromonas gingivalis.
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DOI：
10.1111/omi.12007
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发表时间：
2013-02
影响因子：
3.7
通讯作者：
Lamont RJ
中科院分区：
文献类型：
--
作者：
Hirano T;Beck DA;Wright CJ;Demuth DR;Hackett M;Lamont RJ
牙周病原体牙龈卟啉单胞菌在口腔中经历许多环境条件，并且必须监测和响应各种环境线索。然而，生物体只有五个完整的双组分系统，其中之一是混合系统GppX。为了研究由GppX控制的调节子，我们对ΔgppX突变体进行了RNA-Seq。在ΔgppX突变体中，53个基因表达上调，37个基因表达下调。途径分析显示，在营养充足的条件下，GppX没有系统功能;然而，超过40%的差异丰富的基因被注释为编码假设的蛋白质，表明GppX的新作用。一般来说，GppX的缺失不影响小（s）RNA的缺失。为了进一步确定GppX在调节相对于野生型对照PGN_0151具有最大显著相对丰度变化的假设蛋白质方面的作用，我们构建了一系列菌株，其中ΔgppX突变与含有特异性结构域和磷酸转移突变的GppX蛋白互补。跨膜结构域、DNA结合结构域和磷酸转移残基都是调节PGN_0151所必需的。此外，通过电泳迁移率变动测定（EMSA）证实GppX与PGN_0151启动子区的结合。ΔgppX突变体和Δ PGN_0151突变体均缺乏单种生物膜形成，表明GppX-PGN_0151调节子在生物体的定殖和存活中的作用。
The periodontal pathogen Porphyromonas gingivalis experiences a number of environmental conditions in the oral cavity and must monitor and respond to a variety of environmental cues. However the organism possesses only five full two-component systems, one of which is the hybrid system GppX. To investigate the regulon controlled by GppX we performed RNA-Seq on a ΔgppX mutant. Fifty three genes were up-regulated and 37 genes were down-regulated in the ΔgppX mutant. Pathway analyses revealed no systemic function for GppX under nutrient replete conditions; however, over 40% of the differentially abundant genes were annotated as encoding hypothetical proteins indicating a novel role for GppX. Abundance of small (s)RNA was, in general, not affected by the absence of GppX. To further define the role of GppX with respect to regulation of a hypothetical protein observed with the greatest significant relative abundance change relative to a wild-type control, PGN_0151, we constructed a series of strains in which a ΔgppX mutation was complemented with GppX protein containing specific domain and phosphotransfer mutations. The transmembrane domains, the DNA binding domain and the phosphotransfer residues were all required for regulation of PGN_0151. In addition, binding of GppX to the PGN_0151 promoter regions was confirmed by an electrophoretic mobility shift assay (EMSA). Both the ΔgppX mutant and a ΔPGN_0151 mutant were deficient in monospecies biofilm formation, suggesting a role for the GppX-PGN_0151 regulon in colonization and survival of the organism.
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