Novel stationary-phase-upregulated protein of Porphyromonas gingivalis influences production of superoxide dismutase, thiol peroxidase and thioredoxin

Yuichiro Kikuchi, Naoya Oohara, Keiko Sato, Mamiko Yoshimura, Hideharu Yukitake, Eiko Sakai, Mikio Shoji, Mariko Naito, Koji Nakayama

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Porphyromonas gingivalis, an obligately anaerobic bacterium, is implicated as a major pathogen in the development and progression of chronic periodontitis. Although expression of several virulence factors of the bacterium has been found to be affected by environmental stress such as entrance into the stationary growth phase and heat, there is relatively little information on the mechanisms that may operate in the bacterium in response to environmental stress. In this study, a novel protein (UstA) was investigated that was initially identified following two-dimensional gel analysis. Expression of UstA was upregulated in stationary phase or by exposure to atmospheric oxygen. N-terminal sequencing and database analysis with the P. gingivalis genome sequence revealed that the UstA-encoding gene (ustA) was located upstream of a homologue of the usp gene encoding the universal stress protein on the chromosome. The ustA gene appeared to be transcribed in a monocistronic fashion, as revealed by primer extension and Northern blot analysis. To elucidate the role of UstA in the bacterium, chromosomal mutants carrying a disruption of the ustA gene were constructed. The ustA mutant grew slower than the wild-type parent strain in rich medium, resulting in a lower yield in stationary phase. Furthermore, in this mutant, expression levels of the P. gingivalis homologues of superoxide dismutase, thiol peroxidase and thioredoxin were markedly higher than those in the wild-type, especially in stationary phase. The ustA mutant was more resistant to diamide, a thiol-specific oxidant, than the wild-type. In addition, the ustA mutation suppressed hypersensitivities of the oxyR mutant to diamide, metronidazole and mitomycin C. These results suggest that UstA may play a significant role in oxidative stress responses in the bacterium.

Original languageEnglish
Pages (from-to)841-853
Number of pages13
JournalMicrobiology
Volume151
Issue number3
DOIs
Publication statusPublished - Mar 2005
Externally publishedYes

Fingerprint

Peroxiredoxins
Porphyromonas gingivalis
Sulfhydryl Compounds
Superoxide Dismutase
Diamide
Bacteria
Genes
Proteins
Chronic Periodontitis
Anaerobic Bacteria
Metronidazole
Mitomycin
Virulence Factors
Heat-Shock Proteins
Oxidants
Northern Blotting
Hypersensitivity
Oxidative Stress
Hot Temperature
Chromosomes

ASJC Scopus subject areas

  • Microbiology

Cite this

Novel stationary-phase-upregulated protein of Porphyromonas gingivalis influences production of superoxide dismutase, thiol peroxidase and thioredoxin. / Kikuchi, Yuichiro; Oohara, Naoya; Sato, Keiko; Yoshimura, Mamiko; Yukitake, Hideharu; Sakai, Eiko; Shoji, Mikio; Naito, Mariko; Nakayama, Koji.

In: Microbiology, Vol. 151, No. 3, 03.2005, p. 841-853.

Research output: Contribution to journalArticle

Kikuchi, Yuichiro ; Oohara, Naoya ; Sato, Keiko ; Yoshimura, Mamiko ; Yukitake, Hideharu ; Sakai, Eiko ; Shoji, Mikio ; Naito, Mariko ; Nakayama, Koji. / Novel stationary-phase-upregulated protein of Porphyromonas gingivalis influences production of superoxide dismutase, thiol peroxidase and thioredoxin. In: Microbiology. 2005 ; Vol. 151, No. 3. pp. 841-853.
@article{2647f242d21a49488b0debf9de82b78b,
title = "Novel stationary-phase-upregulated protein of Porphyromonas gingivalis influences production of superoxide dismutase, thiol peroxidase and thioredoxin",
abstract = "Porphyromonas gingivalis, an obligately anaerobic bacterium, is implicated as a major pathogen in the development and progression of chronic periodontitis. Although expression of several virulence factors of the bacterium has been found to be affected by environmental stress such as entrance into the stationary growth phase and heat, there is relatively little information on the mechanisms that may operate in the bacterium in response to environmental stress. In this study, a novel protein (UstA) was investigated that was initially identified following two-dimensional gel analysis. Expression of UstA was upregulated in stationary phase or by exposure to atmospheric oxygen. N-terminal sequencing and database analysis with the P. gingivalis genome sequence revealed that the UstA-encoding gene (ustA) was located upstream of a homologue of the usp gene encoding the universal stress protein on the chromosome. The ustA gene appeared to be transcribed in a monocistronic fashion, as revealed by primer extension and Northern blot analysis. To elucidate the role of UstA in the bacterium, chromosomal mutants carrying a disruption of the ustA gene were constructed. The ustA mutant grew slower than the wild-type parent strain in rich medium, resulting in a lower yield in stationary phase. Furthermore, in this mutant, expression levels of the P. gingivalis homologues of superoxide dismutase, thiol peroxidase and thioredoxin were markedly higher than those in the wild-type, especially in stationary phase. The ustA mutant was more resistant to diamide, a thiol-specific oxidant, than the wild-type. In addition, the ustA mutation suppressed hypersensitivities of the oxyR mutant to diamide, metronidazole and mitomycin C. These results suggest that UstA may play a significant role in oxidative stress responses in the bacterium.",
author = "Yuichiro Kikuchi and Naoya Oohara and Keiko Sato and Mamiko Yoshimura and Hideharu Yukitake and Eiko Sakai and Mikio Shoji and Mariko Naito and Koji Nakayama",
year = "2005",
month = "3",
doi = "10.1099/mic.0.27589-0",
language = "English",
volume = "151",
pages = "841--853",
journal = "Microbiology",
issn = "1350-0872",
publisher = "Society for General Microbiology",
number = "3",

}

TY - JOUR

T1 - Novel stationary-phase-upregulated protein of Porphyromonas gingivalis influences production of superoxide dismutase, thiol peroxidase and thioredoxin

AU - Kikuchi, Yuichiro

AU - Oohara, Naoya

AU - Sato, Keiko

AU - Yoshimura, Mamiko

AU - Yukitake, Hideharu

AU - Sakai, Eiko

AU - Shoji, Mikio

AU - Naito, Mariko

AU - Nakayama, Koji

PY - 2005/3

Y1 - 2005/3

N2 - Porphyromonas gingivalis, an obligately anaerobic bacterium, is implicated as a major pathogen in the development and progression of chronic periodontitis. Although expression of several virulence factors of the bacterium has been found to be affected by environmental stress such as entrance into the stationary growth phase and heat, there is relatively little information on the mechanisms that may operate in the bacterium in response to environmental stress. In this study, a novel protein (UstA) was investigated that was initially identified following two-dimensional gel analysis. Expression of UstA was upregulated in stationary phase or by exposure to atmospheric oxygen. N-terminal sequencing and database analysis with the P. gingivalis genome sequence revealed that the UstA-encoding gene (ustA) was located upstream of a homologue of the usp gene encoding the universal stress protein on the chromosome. The ustA gene appeared to be transcribed in a monocistronic fashion, as revealed by primer extension and Northern blot analysis. To elucidate the role of UstA in the bacterium, chromosomal mutants carrying a disruption of the ustA gene were constructed. The ustA mutant grew slower than the wild-type parent strain in rich medium, resulting in a lower yield in stationary phase. Furthermore, in this mutant, expression levels of the P. gingivalis homologues of superoxide dismutase, thiol peroxidase and thioredoxin were markedly higher than those in the wild-type, especially in stationary phase. The ustA mutant was more resistant to diamide, a thiol-specific oxidant, than the wild-type. In addition, the ustA mutation suppressed hypersensitivities of the oxyR mutant to diamide, metronidazole and mitomycin C. These results suggest that UstA may play a significant role in oxidative stress responses in the bacterium.

AB - Porphyromonas gingivalis, an obligately anaerobic bacterium, is implicated as a major pathogen in the development and progression of chronic periodontitis. Although expression of several virulence factors of the bacterium has been found to be affected by environmental stress such as entrance into the stationary growth phase and heat, there is relatively little information on the mechanisms that may operate in the bacterium in response to environmental stress. In this study, a novel protein (UstA) was investigated that was initially identified following two-dimensional gel analysis. Expression of UstA was upregulated in stationary phase or by exposure to atmospheric oxygen. N-terminal sequencing and database analysis with the P. gingivalis genome sequence revealed that the UstA-encoding gene (ustA) was located upstream of a homologue of the usp gene encoding the universal stress protein on the chromosome. The ustA gene appeared to be transcribed in a monocistronic fashion, as revealed by primer extension and Northern blot analysis. To elucidate the role of UstA in the bacterium, chromosomal mutants carrying a disruption of the ustA gene were constructed. The ustA mutant grew slower than the wild-type parent strain in rich medium, resulting in a lower yield in stationary phase. Furthermore, in this mutant, expression levels of the P. gingivalis homologues of superoxide dismutase, thiol peroxidase and thioredoxin were markedly higher than those in the wild-type, especially in stationary phase. The ustA mutant was more resistant to diamide, a thiol-specific oxidant, than the wild-type. In addition, the ustA mutation suppressed hypersensitivities of the oxyR mutant to diamide, metronidazole and mitomycin C. These results suggest that UstA may play a significant role in oxidative stress responses in the bacterium.

UR - http://www.scopus.com/inward/record.url?scp=15844383444&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=15844383444&partnerID=8YFLogxK

U2 - 10.1099/mic.0.27589-0

DO - 10.1099/mic.0.27589-0

M3 - Article

C2 - 15758230

AN - SCOPUS:15844383444

VL - 151

SP - 841

EP - 853

JO - Microbiology

JF - Microbiology

SN - 1350-0872

IS - 3

ER -