Abstract
Pulsed-field gel electrophoresis (PFGE) analysis revealed that the genomes of some pathogenic Escherichia coli O157:H7 strains, including EDL933, were resistant to NotI digestion. An amino acid sequence comparison suggested that the z2389 gene carried on prophage CP-933R in strain EDL933 is likely to encode a C5-cytosine methyltransferase. The z2389-equivalent gene was found in the NotI-resistant strains tested, but it was not detected in the NotI-susceptible strains. PFGE analysis of the wild-type EDL933 strain and of a z2389 null mutant revealed that z2389 was associated with full genome protection against NotI digestion and partial protection against EagI digestion. In vitro methylation experiments with purified recombinant protein demonstrated that Z2389 is capable of methylating NotI and EagI sites. Sequencing of bisulfite-treated DNA indicated that the methylation occurred at the first cytosine residue of the NotI recognition sequence, whereas EagI sites remained unmethylated or were methylated at the first cytosine residue. Thus, z2389 encodes a DNA cytosine methyltransferase that confers full protection to NotI sites.
| Original language | English |
|---|---|
| Pages (from-to) | 296-303 |
| Number of pages | 8 |
| Journal | International Journal of Medical Microbiology |
| Volume | 300 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 2010 Jun 1 |
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All Science Journal Classification (ASJC) codes
- Microbiology
- Microbiology (medical)
- Infectious Diseases
Cite this
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Identification of prophage gene z2389 in Escherichia coli EDL933 encoding a DNA cytosine methyltransferase for full protection of NotI sites. / Chiou, Chien Shun; Li, Hsin Yi; Tung, Sheng Kai; Chen, Chien Yen; Teng, Ching Hao; Shu, Jwu Ching; Tseng, Joseph T.; Hsu, Chi Yu; Chen, Chien Cheng.
In: International Journal of Medical Microbiology, Vol. 300, No. 5, 01.06.2010, p. 296-303.Research output: Contribution to journal › Article
TY - JOUR
T1 - Identification of prophage gene z2389 in Escherichia coli EDL933 encoding a DNA cytosine methyltransferase for full protection of NotI sites
AU - Chiou, Chien Shun
AU - Li, Hsin Yi
AU - Tung, Sheng Kai
AU - Chen, Chien Yen
AU - Teng, Ching Hao
AU - Shu, Jwu Ching
AU - Tseng, Joseph T.
AU - Hsu, Chi Yu
AU - Chen, Chien Cheng
PY - 2010/6/1
Y1 - 2010/6/1
N2 - Pulsed-field gel electrophoresis (PFGE) analysis revealed that the genomes of some pathogenic Escherichia coli O157:H7 strains, including EDL933, were resistant to NotI digestion. An amino acid sequence comparison suggested that the z2389 gene carried on prophage CP-933R in strain EDL933 is likely to encode a C5-cytosine methyltransferase. The z2389-equivalent gene was found in the NotI-resistant strains tested, but it was not detected in the NotI-susceptible strains. PFGE analysis of the wild-type EDL933 strain and of a z2389 null mutant revealed that z2389 was associated with full genome protection against NotI digestion and partial protection against EagI digestion. In vitro methylation experiments with purified recombinant protein demonstrated that Z2389 is capable of methylating NotI and EagI sites. Sequencing of bisulfite-treated DNA indicated that the methylation occurred at the first cytosine residue of the NotI recognition sequence, whereas EagI sites remained unmethylated or were methylated at the first cytosine residue. Thus, z2389 encodes a DNA cytosine methyltransferase that confers full protection to NotI sites.
AB - Pulsed-field gel electrophoresis (PFGE) analysis revealed that the genomes of some pathogenic Escherichia coli O157:H7 strains, including EDL933, were resistant to NotI digestion. An amino acid sequence comparison suggested that the z2389 gene carried on prophage CP-933R in strain EDL933 is likely to encode a C5-cytosine methyltransferase. The z2389-equivalent gene was found in the NotI-resistant strains tested, but it was not detected in the NotI-susceptible strains. PFGE analysis of the wild-type EDL933 strain and of a z2389 null mutant revealed that z2389 was associated with full genome protection against NotI digestion and partial protection against EagI digestion. In vitro methylation experiments with purified recombinant protein demonstrated that Z2389 is capable of methylating NotI and EagI sites. Sequencing of bisulfite-treated DNA indicated that the methylation occurred at the first cytosine residue of the NotI recognition sequence, whereas EagI sites remained unmethylated or were methylated at the first cytosine residue. Thus, z2389 encodes a DNA cytosine methyltransferase that confers full protection to NotI sites.
UR - http://www.scopus.com/inward/record.url?scp=77952323544&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77952323544&partnerID=8YFLogxK
U2 - 10.1016/j.ijmm.2009.11.003
DO - 10.1016/j.ijmm.2009.11.003
M3 - Article
C2 - 20022807
AN - SCOPUS:77952323544
VL - 300
SP - 296
EP - 303
JO - International Journal of Medical Microbiology
JF - International Journal of Medical Microbiology
SN - 1438-4221
IS - 5
ER -