Identification of prophage gene z2389 in Escherichia coli EDL933 encoding a DNA cytosine methyltransferase for full protection of NotI sites.
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 C(5)-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.
Chiou
Chien-Shun
CS
The Central Region Laboratory, Center of Research and Diagnostics, Centers for Disease Control, Taichung City 40855, Taiwan.
Li
Hsin-Yi
HY
Tung
Sheng-Kai
SK
Chen
Chien-Yen
CY
Teng
Ching-Hao
CH
Shu
Jwu-Ching
JC
Tseng
Joseph T
JT
Hsu
Chi-Yu
CY
Chen
Chien-Cheng
CC
D000818:Animals; D002417:Cattle; D003090:Coliphages / Q000235:genetics*; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics*; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D016521:Electrophoresis, Gel, Pulsed-Field; D019453:Escherichia coli O157 / Q000235:genetics* / Q000302:isolation & purification; D005243:Feces / Q000382:microbiology; D005516:Food Microbiology; D055786:Gene Knockout Techniques; D006801:Humans; D039002:Prophages / Q000201:enzymology* / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics*
D004269:DNA, Bacterial; D014764:Viral Proteins; D004248:DNA (Cytosine-5-)-Methyltransferases; C046751:endodeoxyribonuclease XmaIII; D015252:Deoxyribonucleases, Type II Site-Specific; C038565:GCGGCCGC-specific type II deoxyribonucleases