J Korean Assoc Oral Maxillofac Surg.  2012 Feb;38(1):14-19. 10.5125/jkaoms.2012.38.1.14.

Expression of ssrA in non-pathogen-induced adaptation in the oral cavity through signal exchange with oral pathogens

• Kim SR
• Kwak JW
• Lee SK
• Jung SG
• Han MS
• Kim BS
• Kook MS
• Oh HK
• Park HJ

• KMID: 1434374
• DOI: http://doi.org/10.5125/jkaoms.2012.38.1.14

Abstract

INTRODUCTION
This study was conducted to evaluate ssrA expression resulting from adaptation of Escherichia coli (E. coli) to oral pathogens through signal exchange.
MATERIALS AND METHODS
Human cell lines Hep2 and HT29, wild-type E. coli (WT K-12), ssrA knock-out E. coli (Delta K-12), and Scleropages aureus (S. aureus) were used. A single culture consisting of Hep2, HT29, WT K-12, and Delta K-12, and mixed cultures consisting of Hep2 and WT K-12, Hep2 and Delta K-12, WT K-12 and S. aureus , Delta K-12 and S. aureus , and Hep2, WT K-12, and S. aureus were prepared. For HT29, a mixed culture was prepared with WT K-12 and with WT K-12 and S. aureus. Total RNA was extracted from each culture with the resulting expression of ssrA, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kappaB), and p53 was evaluated by Reverse transcription polymerase chain reaction (RT-PCR).
RESULTS
The expression of ssrA in a single culture of WT K-12 was lower than that observed in the mixed culture of WT K-12 with S. aureus. Greater ssrA expression was observed in the mixed culture of WT K-12 with Hep2 than in the single culture of WT K-12. The expression of NF-kappaB was higher in the mixed culture of Hep2 with Delta K-12 than that in the mixed culture of Hep2 with WT K-12, and was lowest in the single culture of Hep2. The expression of ssrA was higher in the mixed culture of WT K-12 with Hep2 and S. aureus than in the mixed culture of WT K-12 with Hep2.
CONCLUSION
These results suggest that ssrA plays an important role in the mechanism of E. coli adaptation to a new environment.

Keyword

ssrA; Escherichia coli; Adaptation

MeSH Terms

B-Lymphocytes
Cell Line
Escherichia coli
Humans
Mouth
NF-kappa B
Polymerase Chain Reaction
Reverse Transcription
RNA
NF-kappa B
RNA

Figure

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  Fig. 1 Agarose gel fraction of total RNA of K-12 strains stimulated by Scleropages aureus (A). (W: wild-type Escherichia coli [WT K-12], Δ: ssrA knock-out Escherichia coli [Δ K-12], GAPDH: glyceraldehyde 3-phosphate dehydrogenase)

• 

  Fig. 2 Expression of ssrA and GAPDH in single and mixed cultures after stimulation by Scleropages aureus (A). (W: wild-type Escherichia coli [WT K-12], Δ: ssrA knock-out Escherichia coli [Δ K-12], GAPDH: glyceraldehyde 3-phosphate dehydrogenase)

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  Fig. 3 Expression of ssrA in the mixed culture of wild-type Escherichia coli (WT K-12) with Hep2. (Con: control, W: WT K-12, Δ: ssrA knock-out E. coli [Δ K-12], GAPDH: glyceraldehyde 3-phosphate dehydrogenase)

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  Fig. 4 Expression level of NF-κB and p53 from Hep2 and HT29 following K-12 treatment. (C: control, W: wild-type Escherichia coli [WT K-12], Δ: ssrA knock-out E. coli [Δ K-12], GAPDH: glyceraldehyde 3-phosphate dehydrogenase)

• 

  Fig. 5 Expression of ssrA in wild-type Escherichia coli (WT K-12) from a mixed culture with Scleropages aureus (S. aureus). (C: control, W: WT K-12, A: S. aureus, GAPDH: glyceraldehyde 3-phosphate dehydrogenase)

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