PsyR, a transcriptional regulator in quorum sensing system, binds lux box-like sequence in psyI promoter without AHL quorum sensing molecule and activates psyI transcription with AHL in Pseudomonas syringae pv. tabaci 6605

Yuki Ichinose, Yousuke Tasaka, Satoru Yamamoto, Yuko Inoue, Motohiro Takata, Yukiko Nakatsu, Fumiko Taguchi, Mikihiro Yamamoto, Kazuhiro Toyoda, Yoshiteru Noutoshi, Hidenori Matsui

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Abstract

Quorum sensing (QS) is a mechanism for bacterial cell–cell communication using QS signals. N-acyl-homoserine lactones (AHLs), QS signals in Pseudomonas syringae pv. tabaci (Pta) 6605, are synthesized by an AHL synthase (PsyI) and recognized by the cognate transcription factor PsyR. To reveal the role of PsyR in virulence, we generated a ∆psyR mutant and complemented strains of Pta 6605 and found that the ∆psyR mutant is remarkably reduced in AHL production and ability to cause disease and propagate in host tobacco leaves. The phenotypes of complemented strains were restored to that of the wild type (WT). Because the ∆psyR mutant lost nearly all AHL production, we investigated the function of PsyR in the transcription of psyI and production of AHL. Electrophoretic mobility shift assays suggested that the recombinant PsyR protein binds the promoter region of psyI but not psyR without AHL. The addition of AHL did not significantly affect this binding. The binding core sequence of this region was identified as a 20-bp lux box-like sequence. To reveal the function of PsyR and AHL on psyI transcription, we constructed a psyI promoter::lacZYA chimeric reporter gene, and inserted it into the WT and ∆psyI mutant of Pta 6605. β-galactosidase activity increased in a bacterial density-dependent manner in the WT and also in a ∆psyI mutant after the addition of exogenous AHL. These results indicate that the solo PsyR binds the lux box in the psyI promoter and activates transcription in the concomitant presence of AHL.

Original languageEnglish
JournalJournal of General Plant Pathology
DOIs
Publication statusAccepted/In press - Jan 1 2019

Fingerprint

Pseudomonas syringae pv. tabaci
homoserine
quorum sensing
lactones
transcription factors
transcription (genetics)
promoter regions
mutants
galactosidases
recombinant proteins
reporter genes
microbial activity
tobacco
virulence

Keywords

  • AHL
  • PsyI
  • PsyR
  • Quorum sensing

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science

Cite this

@article{fec9559ddf4c41dfb82fa13f67c7e8de,
title = "PsyR, a transcriptional regulator in quorum sensing system, binds lux box-like sequence in psyI promoter without AHL quorum sensing molecule and activates psyI transcription with AHL in Pseudomonas syringae pv. tabaci 6605",
abstract = "Quorum sensing (QS) is a mechanism for bacterial cell–cell communication using QS signals. N-acyl-homoserine lactones (AHLs), QS signals in Pseudomonas syringae pv. tabaci (Pta) 6605, are synthesized by an AHL synthase (PsyI) and recognized by the cognate transcription factor PsyR. To reveal the role of PsyR in virulence, we generated a ∆psyR mutant and complemented strains of Pta 6605 and found that the ∆psyR mutant is remarkably reduced in AHL production and ability to cause disease and propagate in host tobacco leaves. The phenotypes of complemented strains were restored to that of the wild type (WT). Because the ∆psyR mutant lost nearly all AHL production, we investigated the function of PsyR in the transcription of psyI and production of AHL. Electrophoretic mobility shift assays suggested that the recombinant PsyR protein binds the promoter region of psyI but not psyR without AHL. The addition of AHL did not significantly affect this binding. The binding core sequence of this region was identified as a 20-bp lux box-like sequence. To reveal the function of PsyR and AHL on psyI transcription, we constructed a psyI promoter::lacZYA chimeric reporter gene, and inserted it into the WT and ∆psyI mutant of Pta 6605. β-galactosidase activity increased in a bacterial density-dependent manner in the WT and also in a ∆psyI mutant after the addition of exogenous AHL. These results indicate that the solo PsyR binds the lux box in the psyI promoter and activates transcription in the concomitant presence of AHL.",
keywords = "AHL, PsyI, PsyR, Quorum sensing",
author = "Yuki Ichinose and Yousuke Tasaka and Satoru Yamamoto and Yuko Inoue and Motohiro Takata and Yukiko Nakatsu and Fumiko Taguchi and Mikihiro Yamamoto and Kazuhiro Toyoda and Yoshiteru Noutoshi and Hidenori Matsui",
year = "2019",
month = "1",
day = "1",
doi = "10.1007/s10327-019-00893-3",
language = "English",
journal = "Journal of General Plant Pathology",
issn = "1345-2630",
publisher = "Springer Japan",

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TY - JOUR

T1 - PsyR, a transcriptional regulator in quorum sensing system, binds lux box-like sequence in psyI promoter without AHL quorum sensing molecule and activates psyI transcription with AHL in Pseudomonas syringae pv. tabaci 6605

AU - Ichinose, Yuki

AU - Tasaka, Yousuke

AU - Yamamoto, Satoru

AU - Inoue, Yuko

AU - Takata, Motohiro

AU - Nakatsu, Yukiko

AU - Taguchi, Fumiko

AU - Yamamoto, Mikihiro

AU - Toyoda, Kazuhiro

AU - Noutoshi, Yoshiteru

AU - Matsui, Hidenori

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Quorum sensing (QS) is a mechanism for bacterial cell–cell communication using QS signals. N-acyl-homoserine lactones (AHLs), QS signals in Pseudomonas syringae pv. tabaci (Pta) 6605, are synthesized by an AHL synthase (PsyI) and recognized by the cognate transcription factor PsyR. To reveal the role of PsyR in virulence, we generated a ∆psyR mutant and complemented strains of Pta 6605 and found that the ∆psyR mutant is remarkably reduced in AHL production and ability to cause disease and propagate in host tobacco leaves. The phenotypes of complemented strains were restored to that of the wild type (WT). Because the ∆psyR mutant lost nearly all AHL production, we investigated the function of PsyR in the transcription of psyI and production of AHL. Electrophoretic mobility shift assays suggested that the recombinant PsyR protein binds the promoter region of psyI but not psyR without AHL. The addition of AHL did not significantly affect this binding. The binding core sequence of this region was identified as a 20-bp lux box-like sequence. To reveal the function of PsyR and AHL on psyI transcription, we constructed a psyI promoter::lacZYA chimeric reporter gene, and inserted it into the WT and ∆psyI mutant of Pta 6605. β-galactosidase activity increased in a bacterial density-dependent manner in the WT and also in a ∆psyI mutant after the addition of exogenous AHL. These results indicate that the solo PsyR binds the lux box in the psyI promoter and activates transcription in the concomitant presence of AHL.

AB - Quorum sensing (QS) is a mechanism for bacterial cell–cell communication using QS signals. N-acyl-homoserine lactones (AHLs), QS signals in Pseudomonas syringae pv. tabaci (Pta) 6605, are synthesized by an AHL synthase (PsyI) and recognized by the cognate transcription factor PsyR. To reveal the role of PsyR in virulence, we generated a ∆psyR mutant and complemented strains of Pta 6605 and found that the ∆psyR mutant is remarkably reduced in AHL production and ability to cause disease and propagate in host tobacco leaves. The phenotypes of complemented strains were restored to that of the wild type (WT). Because the ∆psyR mutant lost nearly all AHL production, we investigated the function of PsyR in the transcription of psyI and production of AHL. Electrophoretic mobility shift assays suggested that the recombinant PsyR protein binds the promoter region of psyI but not psyR without AHL. The addition of AHL did not significantly affect this binding. The binding core sequence of this region was identified as a 20-bp lux box-like sequence. To reveal the function of PsyR and AHL on psyI transcription, we constructed a psyI promoter::lacZYA chimeric reporter gene, and inserted it into the WT and ∆psyI mutant of Pta 6605. β-galactosidase activity increased in a bacterial density-dependent manner in the WT and also in a ∆psyI mutant after the addition of exogenous AHL. These results indicate that the solo PsyR binds the lux box in the psyI promoter and activates transcription in the concomitant presence of AHL.

KW - AHL

KW - PsyI

KW - PsyR

KW - Quorum sensing

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U2 - 10.1007/s10327-019-00893-3

DO - 10.1007/s10327-019-00893-3

M3 - Article

AN - SCOPUS:85076560127

JO - Journal of General Plant Pathology

JF - Journal of General Plant Pathology

SN - 1345-2630

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