Identification of prophage gene z2389 in Escherichia coli EDL933 encoding a DNA cytosine methyltransferase for full protection of NotI sites

Chien Shun Chiou, Hsin Yi Li, Sheng Kai Tung, Chien Yen Chen, Ching Hao Teng, Jwu Ching Shu, Joseph T. Tseng, Chi Yu Hsu, Chien Cheng Chen

Research output: Contribution to journalArticle

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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 languageEnglish
Pages (from-to)296-303
Number of pages8
JournalInternational Journal of Medical Microbiology
Volume300
Issue number5
DOIs
Publication statusPublished - 2010 Jun 1

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Prophages
Cytosine
Methyltransferases
Escherichia coli
Digestion
Pulsed Field Gel Electrophoresis
DNA
Genes
Genome
Escherichia coli O157
DNA Methylation
Recombinant Proteins
Methylation
Amino Acid Sequence

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Microbiology (medical)
  • Infectious Diseases

Cite this

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. / Identification of prophage gene z2389 in Escherichia coli EDL933 encoding a DNA cytosine methyltransferase for full protection of NotI sites. In: International Journal of Medical Microbiology. 2010 ; Vol. 300, No. 5. pp. 296-303.
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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 journalArticle

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