The sensor histidine kinase PhcS participates in the regulation of quorum sensing-dependent virulence genes in Ralstonia pseudosolanacearum strain OE1-1

ABSTRACT Ralstonia pseudosolanacearum strain OE1-1 secretes methyl 3-hydroxymyristate (3-OH MAME) as a quorum-sensing (QS) signal. Strain OE1-1 senses the chemical by the sensor histidine kinase PhcS, leading to the activation of the LysR family transcriptional regulator PhcA. The activated PhcA con...

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Main Authors: Wakana Senuma, Masayuki Tsuzuki, Chika Takemura, Yuki Terazawa, Akinori Kiba, Kouhei Ohnishi, Kenji Kai, Yasufumi Hikichi
Format: Article
Language:English
Published: American Society for Microbiology 2025-04-01
Series:Microbiology Spectrum
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Online Access:https://journals.asm.org/doi/10.1128/spectrum.00059-25
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author Wakana Senuma
Masayuki Tsuzuki
Chika Takemura
Yuki Terazawa
Akinori Kiba
Kouhei Ohnishi
Kenji Kai
Yasufumi Hikichi
author_facet Wakana Senuma
Masayuki Tsuzuki
Chika Takemura
Yuki Terazawa
Akinori Kiba
Kouhei Ohnishi
Kenji Kai
Yasufumi Hikichi
author_sort Wakana Senuma
collection DOAJ
description ABSTRACT Ralstonia pseudosolanacearum strain OE1-1 secretes methyl 3-hydroxymyristate (3-OH MAME) as a quorum-sensing (QS) signal. Strain OE1-1 senses the chemical by the sensor histidine kinase PhcS, leading to the activation of the LysR family transcriptional regulator PhcA. The activated PhcA controls the expression of QS-dependent genes responsible for QS-regulated phenotypes including virulence. The autophosphorylation of the histidine at amino acid position 230 (H230-PhcS) in PhcS following the 3-OH MAME sensing is required for the PhcA activation. The alternative sensor histidine kinase PhcK is involved in the regulation of phcA, which is independent of 3-OH MAME sensing. Furthermore, the H230Q-PhcS substitution of H230-PhcS with glutamine significantly decreases phcA expression. However, how PhcK and PhcS regulate phcA expression remains unclear. To elucidate the mechanisms of the phcA regulation, we generated a phcK mutant with the H205Q-PhcK substitution of autophosphorylated histidine at amino acid position 205 of PhcK with glutamine. A transcriptome analysis using quantitative real-time polymerase chain reaction assay and RNA sequencing showed that the H230Q-PhcS substitution, but not the H205Q-PhcK substitution, significantly decreased the expression level of phcA. The H230Q-PhcS substitution led to significant changes in the expression levels of QS-dependent genes and a loss of virulence, similar to phcA or phcK deletion. It is thus thought that PhcS participates in not only the 3-OH MAME sensing-independently PhcK-mediated regulation of phcA but also the PhcA activation following 3-OH MAME sensing. Both functions of PhcS are significantly influenced by the autophosphorylation of H230-PhcS.IMPORTANCEThe soil-borne Ralstonia solanacearum species complex (RSSC) infects more than 300 plant species in over 50 families, including solanaceous plants, causing the devastating wilt disease that substantially decreases agricultural production worldwide. The cell density-dependent gene regulation system, QS, is required for RSSC virulence and involves two signaling pathways for the induction and activation of PhcA, which is the master transcriptional regulator in QS. In the present study, we describe the contribution of sensor histidine kinase PhcS to the PhcA induction, along with the alternative sensor kinase PhcK, independently of the sensing of QS signal methyl 3-hydroxymyristate in a phylotype I strain of RSSC, R. pseudosolanacearum strain OE1-1. This study further expands our knowledge of multiple networks, suggesting that several PhcS-mediated two-component systems are likely necessary for RSSC QS and virulence.
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publishDate 2025-04-01
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spelling doaj-art-dcac059fb73f445190f4ffe83471be802025-08-20T02:53:34ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-04-0113410.1128/spectrum.00059-25The sensor histidine kinase PhcS participates in the regulation of quorum sensing-dependent virulence genes in Ralstonia pseudosolanacearum strain OE1-1Wakana Senuma0Masayuki Tsuzuki1Chika Takemura2Yuki Terazawa3Akinori Kiba4Kouhei Ohnishi5Kenji Kai6Yasufumi Hikichi7Faculty of Agriculture and Marine Science, Kochi University, Nankoku, JapanFaculty of Agriculture and Marine Science, Kochi University, Nankoku, JapanFaculty of Agriculture and Marine Science, Kochi University, Nankoku, JapanFaculty of Agriculture and Marine Science, Kochi University, Nankoku, JapanFaculty of Agriculture and Marine Science, Kochi University, Nankoku, JapanFaculty of Agriculture and Marine Science, Kochi University, Nankoku, JapanFaculty of Agriculture and Marine Science, Kochi University, Nankoku, JapanFaculty of Agriculture and Marine Science, Kochi University, Nankoku, JapanABSTRACT Ralstonia pseudosolanacearum strain OE1-1 secretes methyl 3-hydroxymyristate (3-OH MAME) as a quorum-sensing (QS) signal. Strain OE1-1 senses the chemical by the sensor histidine kinase PhcS, leading to the activation of the LysR family transcriptional regulator PhcA. The activated PhcA controls the expression of QS-dependent genes responsible for QS-regulated phenotypes including virulence. The autophosphorylation of the histidine at amino acid position 230 (H230-PhcS) in PhcS following the 3-OH MAME sensing is required for the PhcA activation. The alternative sensor histidine kinase PhcK is involved in the regulation of phcA, which is independent of 3-OH MAME sensing. Furthermore, the H230Q-PhcS substitution of H230-PhcS with glutamine significantly decreases phcA expression. However, how PhcK and PhcS regulate phcA expression remains unclear. To elucidate the mechanisms of the phcA regulation, we generated a phcK mutant with the H205Q-PhcK substitution of autophosphorylated histidine at amino acid position 205 of PhcK with glutamine. A transcriptome analysis using quantitative real-time polymerase chain reaction assay and RNA sequencing showed that the H230Q-PhcS substitution, but not the H205Q-PhcK substitution, significantly decreased the expression level of phcA. The H230Q-PhcS substitution led to significant changes in the expression levels of QS-dependent genes and a loss of virulence, similar to phcA or phcK deletion. It is thus thought that PhcS participates in not only the 3-OH MAME sensing-independently PhcK-mediated regulation of phcA but also the PhcA activation following 3-OH MAME sensing. Both functions of PhcS are significantly influenced by the autophosphorylation of H230-PhcS.IMPORTANCEThe soil-borne Ralstonia solanacearum species complex (RSSC) infects more than 300 plant species in over 50 families, including solanaceous plants, causing the devastating wilt disease that substantially decreases agricultural production worldwide. The cell density-dependent gene regulation system, QS, is required for RSSC virulence and involves two signaling pathways for the induction and activation of PhcA, which is the master transcriptional regulator in QS. In the present study, we describe the contribution of sensor histidine kinase PhcS to the PhcA induction, along with the alternative sensor kinase PhcK, independently of the sensing of QS signal methyl 3-hydroxymyristate in a phylotype I strain of RSSC, R. pseudosolanacearum strain OE1-1. This study further expands our knowledge of multiple networks, suggesting that several PhcS-mediated two-component systems are likely necessary for RSSC QS and virulence.https://journals.asm.org/doi/10.1128/spectrum.00059-25Ralstonia pseudosolanacearumPhcKPhcSquorum sensingvirulence
spellingShingle Wakana Senuma
Masayuki Tsuzuki
Chika Takemura
Yuki Terazawa
Akinori Kiba
Kouhei Ohnishi
Kenji Kai
Yasufumi Hikichi
The sensor histidine kinase PhcS participates in the regulation of quorum sensing-dependent virulence genes in Ralstonia pseudosolanacearum strain OE1-1
Microbiology Spectrum
Ralstonia pseudosolanacearum
PhcK
PhcS
quorum sensing
virulence
title The sensor histidine kinase PhcS participates in the regulation of quorum sensing-dependent virulence genes in Ralstonia pseudosolanacearum strain OE1-1
title_full The sensor histidine kinase PhcS participates in the regulation of quorum sensing-dependent virulence genes in Ralstonia pseudosolanacearum strain OE1-1
title_fullStr The sensor histidine kinase PhcS participates in the regulation of quorum sensing-dependent virulence genes in Ralstonia pseudosolanacearum strain OE1-1
title_full_unstemmed The sensor histidine kinase PhcS participates in the regulation of quorum sensing-dependent virulence genes in Ralstonia pseudosolanacearum strain OE1-1
title_short The sensor histidine kinase PhcS participates in the regulation of quorum sensing-dependent virulence genes in Ralstonia pseudosolanacearum strain OE1-1
title_sort sensor histidine kinase phcs participates in the regulation of quorum sensing dependent virulence genes in ralstonia pseudosolanacearum strain oe1 1
topic Ralstonia pseudosolanacearum
PhcK
PhcS
quorum sensing
virulence
url https://journals.asm.org/doi/10.1128/spectrum.00059-25
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