Comparative functional analysis of the RcsC sensor kinase from different Enterobacteriaceae
Comparative functional analysis of the RcsC sensor kinase from different Enterobacteriaceae
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DOI：
10.1111/j.1574-6968.2009.01543.x
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发表时间：
2009-04-01
影响因子：
2.1
通讯作者：
Clarke, David J.
中科院分区：
文献类型：
--
作者：
Huang, Ya-Hui;Ferrieres, Lionel;Clarke, David J.
Rcs磷接力是整个肠杆菌科中保守的重要信号通路。Rcs磷接力由RcsC传感器激酶、膜定位的含hpt蛋白RcsD和细胞质反应调节因子RcsB组成。在这项研究中，我们感兴趣的是研究不同肠道RcsC同源物之间的功能守恒程度。因此，我们测试了致病性大肠埃希菌、肠炎沙门氏菌和鼠疫耶尔森菌的RcsC同源物对大肠埃希菌K-12中RcsC突变的功能性补充能力。补体表现为：(1)cpsB-lacZ表达的增加，以响应DjlA的过量产生，这是RcsC激活的一个公认的信号；(2)恢复生物膜形成的能力。所有的同源物都增加了cpsB-lacZ的表达，以响应DjlA的过量产生，这证实了不同的RcsC同源物能够感知这种刺激，并将信号转导到大肠杆菌中Rcs途径的下游组分。这表明RcsC的核心信号结构域在整个肠杆菌科中都是功能保守的。然而，我们也发现鼠疫杆菌的RcsC不能在大肠杆菌K-12 RcsC突变体中恢复正常的生物膜形成，我们认为这是由于该同源物观察到的净激酶活性增加。
The Rcs phosphorelay is an important signalling pathway that is conserved throughout the Enterobacteriaceae. The Rcs phosphorelay is composed of the RcsC sensor kinase, a membrane-localized HPt-containing protein RcsD and the cytoplasmic response regulator RcsB. In this study we were interested in studying the degree of functional conservation between the different enteric RcsC homologues. Therefore, we tested for the ability of RcsC homologues from pathogenic Escherichia coli, Salmonella enterica and Yersinia pestis to functionally complement an rcsC mutation in E. coli K-12. Complementation was measured as (1) an increase in cpsB-lacZ expression in response to DjlA overproduction, a well-established signal for RcsC activation and (2) the ability to restore biofilm formation. All of the homologues increased cpsB-lacZ expression in response to DjlA overproduction confirming that the different RcsC homologues are able to sense this stimulus and to transduce the signal to the downstream components of the Rcs pathway in E. coli. This suggests that the core signalling domains of RcsC are functionally conserved throughout the Enterobacteriaceae. However, we also show that RcsC from Y. pestis was unable to restore normal biofilm formation in the E. coli K-12 rcsC mutant and we argue that this is due to the increased net kinase activity observed with this homologue.