Vfr targets promoter of genes encoding methyl-accepting chemotaxis protein in Pseudomonas syringae pv. tabaci 6605.
Vfr targets promoter of genes encoding methyl-accepting chemotaxis protein in Pseudomonas syringae pv. tabaci 6605.
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DOI：
10.1016/j.bbrep.2021.100944
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发表时间：
2021-07
影响因子：
2.7
通讯作者：
Ichinose Y
中科院分区：
文献类型：
--
作者：
Ogura K;Matsui H;Yamamoto M;Noutoshi Y;Toyoda K;Taguchi F;Ichinose Y
毒力因子调节因子（Vfr）是植物病原细菌Pseudomonasscarlingae毒力表达过程中不可缺少的转录因子。然而，Vfr的功能至今还不清楚。vfr的缺失导致了表面群集运动的丧失，并降低了在P. erichingae pv.烟粉虱（Pta）6605.为了鉴定Vfr的靶基因，我们使用密切相关的细菌P.菜B728 a.为此，我们首先产生了具有Pss B728 a中的重组基因vfr：：FLAG的菌株，并使用抗FLAG抗体进行ChIP。纯化免疫沉淀的DNA并用Illumina HiSeq测序。进一步对Vfr：：FLAG特异性峰进行电泳迁移率变化测定，并将PAD_0578、PAD_1776和PAD_2237的基因座标签的启动子区鉴定为Vfr的推定靶基因。这些基因编码植物病原体特异性甲基接受趋化蛋白（Mcp）。这些mcp基因似乎参与了Vfr调控的毒力表达。ChIP-seq和EMSA技术鉴定铜绿假单胞菌Vfr靶基因Vfr靶向3个甲基接受趋化蛋白（mcp）基因。3个MCP基因启动子中存在推定的Vfr结合序列。
Virulence factor regulator (Vfr) is an indispensable transcription factor in the expression of virulence in the phytopathogenic bacteria Pseudomonassyringae. However, the function of Vfr is not known so far. The deletion of vfr resulted in the loss of surface swarming motility and reduced the virulence in P. syringae pv. tabaci (Pta) 6605. In order to identify the target genes of Vfr, we screened the sequences that bind to Vfr by chromatin immune precipitation (ChIP) and sequencing methods using the closely related bacterium P. syringae pv. syringae (Pss) B728a. For this purpose we first generated a strain that possesses the recombinant gene vfr::FLAG in Pss B728a, and performed ChIP using an anti-FLAG antibody. Immunoprecipitated DNA was purified and sequenced with Illumina HiSeq. The Vfr::FLAG-specific peaks were further subjected to an electrophoresis mobility-shift assay, and the promoter regions of locus tag for Psyr_0578 , Psyr_1776, and Psyr_2237 were identified as putative target genes of Vfr. These genes encode plant pathogen–specific methyl-accepting chemotaxis proteins (Mcp). These mcp genes seem to be involved in the Vfr-regulated expression of virulence. Identification of target gene of Vfr in Pseudomonas syringae by ChIP-seq and EMSA. Vfr targets 3 methyl-accepting chemotaxis proteins (mcp) genes. Existence of putative Vfr binding sequences in the promoter of 3 mcp genes.
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