Lrp, a global regulator, regulates the virulence of Vibrio vulnificus

Yu Chi Ho, Feng Ru Hung, Chao Hui Weng, Wei Ting Li, Tzu Hung Chuang, Tsung-lin Liu, Ching Yuan Lin, Chien Jung Lo, Chun Liang Chen, Jen Wei Chen, Masayuki Hashimoto, Lien I. Hor

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Abstract

BACKGROUND: An attenuated mutant (designated NY303) of Vibrio vulnificus, which causes serious wound infection and septicemia in humans, was isolated fortuitously from a clinical strain YJ016. This mutant was defective in cytotoxicity, migration on soft agar and virulence in the mouse. The purpose of this study was to map the mutation in this attenuated mutant and further explore how the gene thus identified is involved in virulence.

METHODS: The whole genome sequence of mutant NY303 determined by next-generation sequencing was compared with that of strain YJ016 to map the mutations. By isolating and characterizing the specific gene-knockout mutants, the gene associated with the phenotype of mutant NY303 was identified. This gene encodes a global regulator, Lrp. A mutant, YH01, deficient in Lrp was isolated and examined in vitro, in vivo and ex vivo to find the affected virulence mechanisms. The target genes of Lrp were further identified by comparing the transcriptomes, which were determined by RNA-seq, of strain YJ016 and mutant YH01. The promoters bound by Lrp were identified by genome footprinting-sequencing, and those related with virulence were further examined by electrophoretic mobility shift assay.

RESULTS: A mutation in lrp was shown to be associated with the reduced cytotoxicity, chemotaxis and virulence of mutant NY303. Mutant YH01 exhibited a phenotype resembling that of mutant NY303, and was defective in colonization in the mouse and growth in mouse serum, but not the antiphagocytosis ability. 596 and 95 genes were down- and up-regulated, respectively, in mutant YH01. Many of the genes involved in secretion of the MARTX cytotoxin, chemotaxis and iron-acquisition were down-regulated in mutant YH01. The lrp gene, which was shown to be negatively autoregulated, and 7 down-regulated virulence-associated genes were bound by Lrp in their promoters. A 14-bp consensus sequence, mkCrTTkwAyTsTG, putatively recognized by Lrp was identified in the promoters of these genes.

CONCLUSIONS: Lrp is a global regulator involved in regulation of cytotoxicity, chemotaxis and iron-acquisition in V. vulnificus. Down-regulation of many of the genes associated with these properties may be responsible, at least partly, for loss of virulence in mutant NY303.

Original languageEnglish
Number of pages1
JournalJournal of biomedical science
Volume24
Issue number1
DOIs
Publication statusPublished - 2017 Aug 11

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Vibrio vulnificus
Virulence
Genes
Chemotaxis
Cytotoxicity
Mutation
Iron
Genome
Phenotype
Gene Knockout Techniques
Aptitude
Cytotoxins
Consensus Sequence
Electrophoretic Mobility Shift Assay
Wound Infection
Transcriptome
Electrophoretic mobility
Agar
Sepsis
Down-Regulation

All Science Journal Classification (ASJC) codes

  • Endocrinology, Diabetes and Metabolism
  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology
  • Biochemistry, medical
  • Pharmacology (medical)

Cite this

Ho, Yu Chi ; Hung, Feng Ru ; Weng, Chao Hui ; Li, Wei Ting ; Chuang, Tzu Hung ; Liu, Tsung-lin ; Lin, Ching Yuan ; Lo, Chien Jung ; Chen, Chun Liang ; Chen, Jen Wei ; Hashimoto, Masayuki ; Hor, Lien I. / Lrp, a global regulator, regulates the virulence of Vibrio vulnificus. In: Journal of biomedical science. 2017 ; Vol. 24, No. 1.
@article{010d029e50c9424eb93731956cb7872d,
title = "Lrp, a global regulator, regulates the virulence of Vibrio vulnificus",
abstract = "BACKGROUND: An attenuated mutant (designated NY303) of Vibrio vulnificus, which causes serious wound infection and septicemia in humans, was isolated fortuitously from a clinical strain YJ016. This mutant was defective in cytotoxicity, migration on soft agar and virulence in the mouse. The purpose of this study was to map the mutation in this attenuated mutant and further explore how the gene thus identified is involved in virulence.METHODS: The whole genome sequence of mutant NY303 determined by next-generation sequencing was compared with that of strain YJ016 to map the mutations. By isolating and characterizing the specific gene-knockout mutants, the gene associated with the phenotype of mutant NY303 was identified. This gene encodes a global regulator, Lrp. A mutant, YH01, deficient in Lrp was isolated and examined in vitro, in vivo and ex vivo to find the affected virulence mechanisms. The target genes of Lrp were further identified by comparing the transcriptomes, which were determined by RNA-seq, of strain YJ016 and mutant YH01. The promoters bound by Lrp were identified by genome footprinting-sequencing, and those related with virulence were further examined by electrophoretic mobility shift assay.RESULTS: A mutation in lrp was shown to be associated with the reduced cytotoxicity, chemotaxis and virulence of mutant NY303. Mutant YH01 exhibited a phenotype resembling that of mutant NY303, and was defective in colonization in the mouse and growth in mouse serum, but not the antiphagocytosis ability. 596 and 95 genes were down- and up-regulated, respectively, in mutant YH01. Many of the genes involved in secretion of the MARTX cytotoxin, chemotaxis and iron-acquisition were down-regulated in mutant YH01. The lrp gene, which was shown to be negatively autoregulated, and 7 down-regulated virulence-associated genes were bound by Lrp in their promoters. A 14-bp consensus sequence, mkCrTTkwAyTsTG, putatively recognized by Lrp was identified in the promoters of these genes.CONCLUSIONS: Lrp is a global regulator involved in regulation of cytotoxicity, chemotaxis and iron-acquisition in V. vulnificus. Down-regulation of many of the genes associated with these properties may be responsible, at least partly, for loss of virulence in mutant NY303.",
author = "Ho, {Yu Chi} and Hung, {Feng Ru} and Weng, {Chao Hui} and Li, {Wei Ting} and Chuang, {Tzu Hung} and Tsung-lin Liu and Lin, {Ching Yuan} and Lo, {Chien Jung} and Chen, {Chun Liang} and Chen, {Jen Wei} and Masayuki Hashimoto and Hor, {Lien I.}",
year = "2017",
month = "8",
day = "11",
doi = "10.1186/s12929-017-0361-9",
language = "English",
volume = "24",
journal = "Journal of Biomedical Science",
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}

Ho, YC, Hung, FR, Weng, CH, Li, WT, Chuang, TH, Liu, T, Lin, CY, Lo, CJ, Chen, CL, Chen, JW, Hashimoto, M & Hor, LI 2017, 'Lrp, a global regulator, regulates the virulence of Vibrio vulnificus', Journal of biomedical science, vol. 24, no. 1. https://doi.org/10.1186/s12929-017-0361-9

Lrp, a global regulator, regulates the virulence of Vibrio vulnificus. / Ho, Yu Chi; Hung, Feng Ru; Weng, Chao Hui; Li, Wei Ting; Chuang, Tzu Hung; Liu, Tsung-lin; Lin, Ching Yuan; Lo, Chien Jung; Chen, Chun Liang; Chen, Jen Wei; Hashimoto, Masayuki; Hor, Lien I.

In: Journal of biomedical science, Vol. 24, No. 1, 11.08.2017.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Lrp, a global regulator, regulates the virulence of Vibrio vulnificus

AU - Ho, Yu Chi

AU - Hung, Feng Ru

AU - Weng, Chao Hui

AU - Li, Wei Ting

AU - Chuang, Tzu Hung

AU - Liu, Tsung-lin

AU - Lin, Ching Yuan

AU - Lo, Chien Jung

AU - Chen, Chun Liang

AU - Chen, Jen Wei

AU - Hashimoto, Masayuki

AU - Hor, Lien I.

PY - 2017/8/11

Y1 - 2017/8/11

N2 - BACKGROUND: An attenuated mutant (designated NY303) of Vibrio vulnificus, which causes serious wound infection and septicemia in humans, was isolated fortuitously from a clinical strain YJ016. This mutant was defective in cytotoxicity, migration on soft agar and virulence in the mouse. The purpose of this study was to map the mutation in this attenuated mutant and further explore how the gene thus identified is involved in virulence.METHODS: The whole genome sequence of mutant NY303 determined by next-generation sequencing was compared with that of strain YJ016 to map the mutations. By isolating and characterizing the specific gene-knockout mutants, the gene associated with the phenotype of mutant NY303 was identified. This gene encodes a global regulator, Lrp. A mutant, YH01, deficient in Lrp was isolated and examined in vitro, in vivo and ex vivo to find the affected virulence mechanisms. The target genes of Lrp were further identified by comparing the transcriptomes, which were determined by RNA-seq, of strain YJ016 and mutant YH01. The promoters bound by Lrp were identified by genome footprinting-sequencing, and those related with virulence were further examined by electrophoretic mobility shift assay.RESULTS: A mutation in lrp was shown to be associated with the reduced cytotoxicity, chemotaxis and virulence of mutant NY303. Mutant YH01 exhibited a phenotype resembling that of mutant NY303, and was defective in colonization in the mouse and growth in mouse serum, but not the antiphagocytosis ability. 596 and 95 genes were down- and up-regulated, respectively, in mutant YH01. Many of the genes involved in secretion of the MARTX cytotoxin, chemotaxis and iron-acquisition were down-regulated in mutant YH01. The lrp gene, which was shown to be negatively autoregulated, and 7 down-regulated virulence-associated genes were bound by Lrp in their promoters. A 14-bp consensus sequence, mkCrTTkwAyTsTG, putatively recognized by Lrp was identified in the promoters of these genes.CONCLUSIONS: Lrp is a global regulator involved in regulation of cytotoxicity, chemotaxis and iron-acquisition in V. vulnificus. Down-regulation of many of the genes associated with these properties may be responsible, at least partly, for loss of virulence in mutant NY303.

AB - BACKGROUND: An attenuated mutant (designated NY303) of Vibrio vulnificus, which causes serious wound infection and septicemia in humans, was isolated fortuitously from a clinical strain YJ016. This mutant was defective in cytotoxicity, migration on soft agar and virulence in the mouse. The purpose of this study was to map the mutation in this attenuated mutant and further explore how the gene thus identified is involved in virulence.METHODS: The whole genome sequence of mutant NY303 determined by next-generation sequencing was compared with that of strain YJ016 to map the mutations. By isolating and characterizing the specific gene-knockout mutants, the gene associated with the phenotype of mutant NY303 was identified. This gene encodes a global regulator, Lrp. A mutant, YH01, deficient in Lrp was isolated and examined in vitro, in vivo and ex vivo to find the affected virulence mechanisms. The target genes of Lrp were further identified by comparing the transcriptomes, which were determined by RNA-seq, of strain YJ016 and mutant YH01. The promoters bound by Lrp were identified by genome footprinting-sequencing, and those related with virulence were further examined by electrophoretic mobility shift assay.RESULTS: A mutation in lrp was shown to be associated with the reduced cytotoxicity, chemotaxis and virulence of mutant NY303. Mutant YH01 exhibited a phenotype resembling that of mutant NY303, and was defective in colonization in the mouse and growth in mouse serum, but not the antiphagocytosis ability. 596 and 95 genes were down- and up-regulated, respectively, in mutant YH01. Many of the genes involved in secretion of the MARTX cytotoxin, chemotaxis and iron-acquisition were down-regulated in mutant YH01. The lrp gene, which was shown to be negatively autoregulated, and 7 down-regulated virulence-associated genes were bound by Lrp in their promoters. A 14-bp consensus sequence, mkCrTTkwAyTsTG, putatively recognized by Lrp was identified in the promoters of these genes.CONCLUSIONS: Lrp is a global regulator involved in regulation of cytotoxicity, chemotaxis and iron-acquisition in V. vulnificus. Down-regulation of many of the genes associated with these properties may be responsible, at least partly, for loss of virulence in mutant NY303.

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