Dataset Information

Analysis of HrpG regulons and HrpG-interacting proteins by ChIP-seq and affinity proteomics in Xanthomonas campestris.

ABSTRACT: Gamma-proteobacteria Xanthomonas spp. cause at least 350 different plant diseases among important agricultural crops, which result in serious yield losses. Xanthomonas spp. 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, and hrpE, 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 of Xanthomonas campestris pv. campestris (HU_xcc ) was proved to be involved in bacterial virulence by increasing the complexity and intelligence of the bacterial signalling pathways in the T3SS.

SUBMITTER: Zhang HY

PROVIDER: S-EPMC7036363 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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Publications

Analysis of HrpG regulons and HrpG-interacting proteins by ChIP-seq and affinity proteomics in Xanthomonas campestris.

Zhang Hong-Yu HY Wei Jin-Wei JW Qian Wei W Deng Chao-Ying CY

Molecular plant pathology 20200108 3

Gamma-proteobacteria Xanthomonas spp. cause at least 350 different plant diseases among important agricultural crops, which result in serious yield losses. Xanthomonas spp. 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 purifica ...[more]

PMID: 31916392

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Project description:Xanthomonas axonopodis pv. citri (XAC) is the causal agent of citrus canker which is one of the most serious diseases of citrus. To understand the virulence mechanism of XAC, we designed and conducted genome-wide microarray analyses to characterize the regulons of HrpG and HrpX, which are critical for the pathogenicity of XAC. Our analyses revealed that the expression of 232 genes and 181 genes belonged to the HrpG and HrpX regulons, respectively. Totally, 123 genes were overlapped in the two regulons at any of the three time-points. Our results showed that HrpG and HrpX regulated all 25 T3SS genes, 22 T3SS effector genes, and 29 T2SS substrate genes. Our data showed that XAC regulates multiple cellular activities responding to the host environment, such as amino acid biosynthesis, oxidative phosphorylation, pentose-phosphate pathway, transport of sugar, iron and potassium, the phenolic compounds catabolism pathway (pcaQ) through HrpX and HrpG. We also identified 124 and 90 unknown genes controlled by HrpG and HrpX, respectively. Our data suggest that a cross-talk exists between HrpG and quorum sensing system in XAC. We also identified genes involved in chemotaxis and flagellar biosynthesis, production of extracellular enzymes, metabolic pathways, as well as signal transduction controlled by HrpG/HrpX. Our results suggest that HrpG and HrpX interplay with global signaling network and co-ordinate the expression of multiple virulence factors for modification and adaption of host environment during XAC infection. Two mutants and three time-points experiment. Four biological and dye-swap replicates of each strain per each time-point were used. Totally, there were three datasets of hrpG mutant: 1) hrpG mutant vs. wild type strain at time-point A; 2) hrpG mutant vs. wild type strain at time-point B; 3) hrpG mutant vs. wild type strain at time-point C. There are also three datasets of hrpX mutant: 1) hrpX mutant vs. wild type strain at time-point A; 2) hrpX mutant vs. wild type strain at time-point B; 3) hrpX mutant vs. wild type strain at time-point C.

Dataset Information

Analysis of HrpG regulons and HrpG-interacting proteins by ChIP-seq and affinity proteomics in Xanthomonas campestris.

Publications

Analysis of HrpG regulons and HrpG-interacting proteins by ChIP-seq and affinity proteomics in Xanthomonas campestris.

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