Identification and characterization of PhoP regulon members in Yersinia pestis biovar Microtus.
Identification and characterization of PhoP regulon members in Yersinia pestis biovar Microtus.
复制标题
鼠疫耶尔森氏菌 Biovar Microtus 中 PhoP 调节子成员的鉴定和表征。
DOI：
10.1186/1471-2164-9-143
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发表时间：
2008-03-27
期刊：
影响因子：
4.4
通讯作者：
Yang, Ruifu
中科院分区：
文献类型：
--
作者：
Li, Yingli;Gao, He;Qin, Long;Li, Bei;Han, Yanping;Guo, Zhaobiao;Song, Yajun;Zhai, Junhui;Du, Zongmin;Wang, Xiaoyi;Zhou, Dongsheng;Yang, Ruifu
转录调节因子PhoP已被证明对鼠疫杆菌在巨噬细胞和各种体外应激下的存活很重要。然而，PhoP促进细菌胞内存活的机制尚不完全清楚。我们之前的微阵列分析表明，PhoP在鼠疫杆菌中控制着广泛的细胞途径。本文通过一系列的生化实验研究了鼠疫菌生物变种田鼠PhoP调控的成员。通过凝胶迁移量转移实验和定量RT-PCR，共有30个推测的转录单位被表征为PhoP的直接靶点。引物延伸试验进一步用于确定18个php依赖启动子的转录起始位点，并定位-10和-35元件。DNase I足迹用于鉴定17个PhoP依赖启动子中的PhoP结合位点，从而鉴定出PhoP盒和矩阵，它们都代表了鼠疫菌PhoP识别的保守信号。本文提供的数据为模拟phop启动子DNA相互作用提供了良好的基础，这对phop介导的转录调控至关重要。已证实的PhoP直接靶点包括9个编码调控因子的基因和21个具有解毒、DNA损伤保护、抗抗菌肽和镁限制适应功能的基因或操纵子。我们可以推测PhoP是一个全球性的调控因子，它不仅通过直接控制特定基因的表达，还通过作用于一组专门的调控因子间接调节各种细胞通路，从而控制复杂的调控级联。这些结果有助于我们深入了解鼠疫杆菌在宿主巨噬细胞采用的抗菌策略下存活的php依赖机制。
The transcription regulator PhoP has been shown to be important for Y. pestis survival in macrophages and under various in vitro stresses. However, the mechanism by which PhoP promotes bacterial intracellular survival is not fully understood. Our previous microarray analysis suggested that PhoP governed a wide set of cellular pathways in Y. pestis. A series of biochemical experiments were done herein to study members of the PhoP regulon of Y. pestis biovar Microtus. By using gel mobility shift assay and quantitative RT-PCR, a total of 30 putative transcription units were characterized as direct PhoP targets. The primer extension assay was further used to determine the transcription start sites of 18 PhoP-dependent promoters and to localize the -10 and -35 elements. The DNase I footprinting was used to identify the PhoP-binding sites within 17 PhoP-dependent promoters, enabling the identification of PhoP box and matrix that both represented the conserved signals for PhoP recognition in Y. pestis. Data presented here providing a good basis for modeling PhoP-promoter DNA interactions that is crucial to the PhoP-mediated transcriptional regulation. The proven direct PhoP targets include nine genes encoding regulators and 21 genes or operons with functions of detoxification, protection against DNA damages, resistance to antimicrobial peptides, and adaptation to magnesium limitation. We can presume that PhoP is a global regulator that controls a complex regulatory cascade by a mechanism of not only directly controlling the expression of specific genes, but also indirectly regulating various cellular pathways by acting on a set of dedicated regulators. These results help us gain insights into the PhoP-dependent mechanisms by which Y. pestis survives the antibacterial strategies employed by host macrophages.
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影响因子：
9.5
作者：
Olson, Sue A
通讯作者：
Olson, Sue A
DOI：
10.1073/pnas.120163297
发表时间：
2000-06-06
影响因子：
11.1
作者：
Datsenko, KA;Wanner, BL
通讯作者：
Wanner, BL
DOI：
10.1073/pnas.0408026101
发表时间：
2004-12-21
影响因子：
11.1
作者：
Achtman, M;Morelli, G;Keim, P
通讯作者：
Keim, P
影响因子：
3.2
作者：
Lejona, S;Aguirre, A;Soncini, FC
通讯作者：
Soncini, FC
影响因子：
3.9
作者：
Monsieurs, P;De Keersmaecker, S;Marchal, K
通讯作者：
Marchal, K