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Identification of a Campylobacter coli methyltransferase targeting adenines at GATC sites
Vikrant Dutta1, Eric Altermann2,3, Maria D Crespo1
1Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC 27695, USA.
Abstract:
Campylobacter coli can infect humans and colonize multiple other animals, but its host-associated genes or adaptations are poorly understood. Adenine methylation at GATC sites, resulting in MboI resistance of genomic DNA, was earlier frequently detected among C. coli from swine but not among turkey-derived isolates. The underlying genetic basis has remained unknown. Comparative genome sequence analyses of C. coli 6461, a swine-derived strain with MboI-resistant DNA, revealed two chromosomal ORFs, 0059 and 0060, encoding a putative DNA methyltransferase and a conserved hypothetical protein, respectively, which were lacking from the genome of the turkey-derived C. coli strain 11601, which had MboI-susceptible DNA. To determine whether ORF0059 mediated MboI resistance and hence encoded a putative N6-adenine DNA methyltransferase, the gene was cloned immediately upstream of a chloramphenicol resistance cassette (cat) and a PCR fragment harboring ORF0059-cat was transformed into C. coli 11601. The transformants had MboI-resistant DNA, suggesting a direct role of this gene in methylation of adenines at GATC sites. In silico analyses suggested that the ORF0059-ORF0060 cassette was more frequent among C. coli from swine than certain other sources (e.g. cattle, humans). Potential impacts of ORF0059-mediated methylation on C. coli host preference and other adaptations remain to be elucidated.
Insights
A novel DNA methyltransferase gene (ORF0059) in Campylobacter coli confers resistance to MboI restriction by methylating adenine at GATC sites. This finding explains differences in MboI resistance between swine and turkey strains.
Area of Science:
- Microbiology
- Genomics
- Molecular Biology
Background:
- Campylobacter coli infects humans and animals, but its host-specific adaptations are poorly understood.
- Adenine methylation at GATC sites confers MboI resistance, observed in swine-derived C. coli but not turkey-derived strains.
Purpose of the Study:
- To identify the genetic basis for MboI resistance in Campylobacter coli.
- To investigate the role of specific genes in mediating adenine methylation and MboI resistance.
Main Methods:
- Comparative genome sequencing of swine and turkey C. coli strains.
- Gene cloning and transformation experiments to assess functional roles.
- In silico analyses to determine gene frequency across different host sources.
Main Results:
- Two open reading frames (ORFs), 0059 and 0060, were identified in a MboI-resistant swine strain, absent in a susceptible turkey strain.
- Cloning and transformation confirmed that ORF0059 encodes an N6-adenine DNA methyltransferase responsible for MboI resistance.
- In silico analysis indicated the ORF0059-ORF0060 cassette is more prevalent in swine-associated C. coli.
Conclusions:
- ORF0059 is the key gene responsible for adenine methylation at GATC sites, conferring MboI resistance in Campylobacter coli.
- The presence of this methylation system may influence C. coli's host preference and adaptation strategies.
- Further research is needed to elucidate the full impact of ORF0059-mediated methylation on C. coli biology.
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