HrpS Is a Global Regulator on Type III Secretion System (T3SS) and Non-T3SS Genes in Pseudomonas savastanoi pv. phaseolicola

0301 basic medicine Chromatin Immunoprecipitation Botany High-Throughput Nucleotide Sequencing Electrophoretic Mobility Shift Assay Gene Expression Regulation, Bacterial Sequence Analysis, DNA Microbiology Models, Biological QR1-502 DNA-Binding Proteins 03 medical and health sciences Bacterial Proteins Genes, Reporter QK1-989 Biofilms Pseudomonas Type III Secretion Systems Point Mutation Nucleotide Motifs Promoter Regions, Genetic Transcription Factors
DOI: 10.1094/mpmi-02-18-0035-r Publication Date: 2018-05-02T15:38:09Z
ABSTRACT
The type III secretion system (T3SS) is the main machinery for Pseudomonas savastanoi and other gram-negative bacteria to invade plant cells. HrpR and HrpS form a hetero-hexamer, which activates the expression of HrpL, which induces all T3SS genes by binding to a ‘hrp box’ in promoters. However, the individual molecular mechanism of HrpR or HrpS has not been fully understood. Through chromatin immunoprecipitation coupled to high-throughput DNA sequencing, we found that HrpR, HrpS, and HrpL had four, 47, and 31 targets on the genome, respectively. HrpS directly bound to the promoter regions of a group of T3SS genes and non-T3SS genes. HrpS independently regulated these genes in a hrpL deletion strain. Additionally, a HrpS-binding motif (GTGCCAAA) was identified, which was verified by electrophoretic mobility shift assay and lux-reporter assay. HrpS also regulated motility and biofilm formation in P. savastanoi. The present study strongly suggests that HrpS alone can work as a global regulator on both T3SS and non-T3SS genes in P. savastanoi. [Formula: see text] Copyright © 2018 The Author(s). This is an open-access article distributed under the CC BY-NC-ND 4.0 International license .
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