Abstract
Aggregatibacter actinomycetemcomitans, a potent pathogen of periodontitis, typically grows as a rough and adherent colony on primary isolated cultures. The colony transforms into a smooth phenotype during repeated subculture. In this study, we aimed to identify highly expressed genes in the rough-colony-forming phenotype for isolation of host-induced genes. Using a cDNA-subtractive hybridization technique, three genes, homologous to a macrophage infectivity potentiator gene (mip), peroxiredoxin gene (prx) and outer membrane protein gene (ompA), were identified. The expression levels of these genes in the rough-colony-forming phenotype were 4-10-fold higher as compared with the smooth-colony-forming phenotype. Attention was focused on the mip-like gene, and a recombinant protein and a deficient mutant were constructed. The recombinant protein reacted with sera from patients with periodontitis, suggesting the production of the Mip-like protein in periodontal lesions. Viable quantitative invasion assay demonstrated that the viable cell counts of the wild-type strain that invaded HeLa cells were more than fourfold as compared with the mip-deficient mutant. The expression of the mip-like gene, prx-like gene and ompA-like gene may be enhanced in the host, and the mip-like gene may play an important role in the infection of A. actinomycetemcomitans, especially in its invasion of the epithelium.
| Original language | English |
|---|---|
| Pages (from-to) | 226-236 |
| Number of pages | 11 |
| Journal | FEMS Immunology and Medical Microbiology |
| Volume | 58 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 2010 |
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Keywords
- Aggregatibacter actinomycetemcomitans
- Host-induced gene
- Invasion
- Macrophage infectivity potentiator (mip)
- Rough-colony-forming phenotype
ASJC Scopus subject areas
- Immunology
- Immunology and Allergy
- Infectious Diseases
- Microbiology (medical)
- Microbiology
- Medicine(all)
Cite this
Highly expressed genes in a rough-colony-forming phenotype of Aggregatibacter actinomycetemcomitans : Implication of a mip-like gene for the invasion of host tissue. / Maeda, Takemasa; Maeda, Hiroshi; Yamabe, Kokoro; Mineshiba, Junji; Tanimoto, Ichiro; Yamamoto, Tadashi; Naruishi, Koji; Kokeguchi, Susumu; Takashiba, Shogo.
In: FEMS Immunology and Medical Microbiology, Vol. 58, No. 2, 2010, p. 226-236.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Highly expressed genes in a rough-colony-forming phenotype of Aggregatibacter actinomycetemcomitans
T2 - Implication of a mip-like gene for the invasion of host tissue
AU - Maeda, Takemasa
AU - Maeda, Hiroshi
AU - Yamabe, Kokoro
AU - Mineshiba, Junji
AU - Tanimoto, Ichiro
AU - Yamamoto, Tadashi
AU - Naruishi, Koji
AU - Kokeguchi, Susumu
AU - Takashiba, Shogo
PY - 2010
Y1 - 2010
N2 - Aggregatibacter actinomycetemcomitans, a potent pathogen of periodontitis, typically grows as a rough and adherent colony on primary isolated cultures. The colony transforms into a smooth phenotype during repeated subculture. In this study, we aimed to identify highly expressed genes in the rough-colony-forming phenotype for isolation of host-induced genes. Using a cDNA-subtractive hybridization technique, three genes, homologous to a macrophage infectivity potentiator gene (mip), peroxiredoxin gene (prx) and outer membrane protein gene (ompA), were identified. The expression levels of these genes in the rough-colony-forming phenotype were 4-10-fold higher as compared with the smooth-colony-forming phenotype. Attention was focused on the mip-like gene, and a recombinant protein and a deficient mutant were constructed. The recombinant protein reacted with sera from patients with periodontitis, suggesting the production of the Mip-like protein in periodontal lesions. Viable quantitative invasion assay demonstrated that the viable cell counts of the wild-type strain that invaded HeLa cells were more than fourfold as compared with the mip-deficient mutant. The expression of the mip-like gene, prx-like gene and ompA-like gene may be enhanced in the host, and the mip-like gene may play an important role in the infection of A. actinomycetemcomitans, especially in its invasion of the epithelium.
AB - Aggregatibacter actinomycetemcomitans, a potent pathogen of periodontitis, typically grows as a rough and adherent colony on primary isolated cultures. The colony transforms into a smooth phenotype during repeated subculture. In this study, we aimed to identify highly expressed genes in the rough-colony-forming phenotype for isolation of host-induced genes. Using a cDNA-subtractive hybridization technique, three genes, homologous to a macrophage infectivity potentiator gene (mip), peroxiredoxin gene (prx) and outer membrane protein gene (ompA), were identified. The expression levels of these genes in the rough-colony-forming phenotype were 4-10-fold higher as compared with the smooth-colony-forming phenotype. Attention was focused on the mip-like gene, and a recombinant protein and a deficient mutant were constructed. The recombinant protein reacted with sera from patients with periodontitis, suggesting the production of the Mip-like protein in periodontal lesions. Viable quantitative invasion assay demonstrated that the viable cell counts of the wild-type strain that invaded HeLa cells were more than fourfold as compared with the mip-deficient mutant. The expression of the mip-like gene, prx-like gene and ompA-like gene may be enhanced in the host, and the mip-like gene may play an important role in the infection of A. actinomycetemcomitans, especially in its invasion of the epithelium.
KW - Aggregatibacter actinomycetemcomitans
KW - Host-induced gene
KW - Invasion
KW - Macrophage infectivity potentiator (mip)
KW - Rough-colony-forming phenotype
UR - http://www.scopus.com/inward/record.url?scp=76349084700&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=76349084700&partnerID=8YFLogxK
U2 - 10.1111/j.1574-695X.2009.00624.x
DO - 10.1111/j.1574-695X.2009.00624.x
M3 - Article
C2 - 19903198
AN - SCOPUS:76349084700
VL - 58
SP - 226
EP - 236
JO - Pathogens and Disease
JF - Pathogens and Disease
SN - 2049-632X
IS - 2
ER -