Analysis of HrpG regulons and HrpG-interacting proteins by ChIP-seq and affinity proteomics in Xanthomonas campestris
Analysis of HrpG regulons and HrpG-interacting proteins by ChIP-seq and affinity proteomics in Xanthomonas campestris
复制标题
通过 ChIP-seq 和亲和蛋白质组学分析野油菜黄单胞菌中的 HrpG 调节子和 HrpG 相互作用蛋白
DOI：
10.1111/mpp.12903
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发表时间：
2020
影响因子：
4.9
通讯作者：
Deng C. Y.
中科院分区：
文献类型：
--
作者：
Zhang H. Y;Wei J. W;Qian W;Deng C. Y.
γproteobacteriaXanthomonasspp。在重要的农作物中造成至少350种不同的植物病害，造成严重的产量损失。主要依靠III型分泌系统（T3SS）感染宿主并在非宿主中诱导过敏反应。HrpG是T3SS的主要调控因子，在细菌毒力中起主导作用。在这项研究中，我们使用染色质免疫沉淀后测序（ChIP‐seq）和串联亲和纯化（TAP）来系统地表征HrpG调控子和HrpG相互作用蛋白。我们从ChIP - seq分析中获得186个候选HrpG下游基因，这些基因代表了全基因组范围的调控谱。获得了一个一致的HrpG结合基序，并证实了三个T3SS基因hpa2、hrcU和hrpe在诱导培养基中被HrpG直接转录激活。从TAP数据中共鉴定出273个推测的HrpG相互作用蛋白，并且通过增加T3SS中细菌信号通路的复杂性和智能，证明了campestris（HUxcc）的DNA结合组蛋白样HU蛋白参与细菌毒力。
Gamma‐proteobacteriaXanthomonasspp. cause at least 350 different plant diseases among important agricultural crops, which result in serious yield losses.Xanthomonasspp. rely mainly on the type III secretion system (T3SS) to infect their hosts and induce a hypersensitive response in nonhosts. HrpG, the master regulator of the T3SS, plays the dominant role in bacterial virulence. In this study, we used chromatin immunoprecipitation followed by sequencing (ChIP‐seq) and tandem affinity purification (TAP) to systematically characterize the HrpG regulon and HrpG interacting proteins in vivo. We obtained 186 candidate HrpG downstream genes from the ChIP‐seq analysis, which represented the genomic‐wide regulon spectrum. A consensus HrpG‐binding motif was obtained and three T3SS genes,hpa2,hrcU, andhrpE, were confirmed to be directly transcriptionally activated by HrpG in the inducing medium. A total of 273 putative HrpG interacting proteins were identified from the TAP data and the DNA‐binding histone‐like HU protein ofXanthomonas campestrispv.campestris(HUxcc) was proved to be involved in bacterial virulence by increasing the complexity and intelligence of the bacterial signalling pathways in the T3SS.