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. 2021 Oct;7(10):000679.
doi: 10.1099/mgen.0.000679.

Genome and Methylome analysis of a phylogenetic novel Campylobacter coli cluster with C. jejuni introgression

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Genome and Methylome analysis of a phylogenetic novel Campylobacter coli cluster with C. jejuni introgression

Anastasia-Lisa Dieckmann et al. Microb Genom. 2021 Oct.

Abstract

The intriguing recent discovery of Campylobacter coli strains, especially of clade 1, that (i) possess mosaic C. coli/C. jejuni alleles, (ii) demonstrate mixed multilocus sequence types (MLSTs) and (iii) have undergone genome-wide introgression has led to the speculation that these two species may be involved in an accelerated rate of horizontal gene transfer that is progressively leading to the merging of both species in a process coined 'despeciation'. In an MLST-based neighbour-joining tree of a number of C. coli and C. jejuni isolates of different clades, three prominent Campylobacter isolates formed a seemingly separate cluster besides the previously described C. coli and C. jejuni clades. In the light of the suspected, ongoing genetic introgression between the C. coli and C. jejuni species, this cluster of Campylobacter isolates is proposed to present one of the hybrid clonal complexes in the despeciation process of the genus. Specific DNA methylation as well as restriction modification systems are known to be involved in selective uptake of external DNA and their role in such genetic introgression remains to be further investigated. In this study, the phylogeny and DNA methylation of these putative C. coli/C. jejuni hybrid strains were explored, their genomic mosaic structure caused by C. jejuni introgression was demonstrated and basic phenotypic assays were used to characterize these isolates. The genomes of the three hybrid Campylobacter strains were sequenced using PacBio SMRT sequencing, followed by methylome analysis by Restriction-Modification Finder and genome analysis by Parsnp, Smash++ and blast. Additionally, the strains were phenotypically characterized with respect to growth behaviour, motility, eukaryotic cell invasion and adhesion, autoagglutination, biofilm formation, and water survival ability. Our analyses show that the three hybrid Campylobacter strains are clade 1 C. coli strains, which have acquired between 8.1 and 9.1 % of their genome from C. jejuni. The C. jejuni genomic segments acquired are distributed over the entire genome and do not form a coherent cluster. Most of the genes originating from C. jejuni are involved in chemotaxis and motility, membrane transport, cell signalling, or the resistance to toxic compounds such as bile acids. Interspecies gene transfer from C. jejuni has contributed 8.1-9.1% to the genome of three C. coli isolates and initiated the despeciation between C. jejuni and C. coli. Based on their functional annotation, the genes originating from C. jejuni enable the adaptation of the three strains to an intra-intestinal habitat. The transfer of a fused type II restriction-modification system that recognizes the CAYNNNNNCTC/GAGNNNNNRTG motif seems to be the key for the recombination of the C. jejuni genetic material with C. coli genomes.

Keywords: Campylobacter coli; Campylobacter jejuni; PacBio SMRT sequencing; genome; introgression; methylome.

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Conflict of interest statement

The authors declare that there are no conflicts of interest.

Figures

Fig. 1.
Fig. 1.
MLST-based phylogenetic tree of C. coli and C. jejuni . MLST-based dendrogram including sequences of 41 strains places the three hybrid strains (red) between C. jejuni (yellow) and C. coli (clades 1A green, 1B dark green, 1C pale green, clade 2 purple, clade 3 blue).
Fig. 2.
Fig. 2.
Core genome analysis of representative genomes of C. coli and C. jejuni by Parsnp. The core genome-based analysis of 20 C . coli and four C . jejuni genomes identifies the isolates meC0280, meC0281 and meC0467 as C. coli clade 1 representatives.
Fig. 3.
Fig. 3.
Similarity analysis of hybrid strains to C. coli or C. jejuni references. Genome segments of 1000 bp (Smash++ algorithm) are matched to the genomes of C. coli BfR-CA-9557 and C. jejuni NCTC 11168. (a) Segments that are more similar to C. coli are shown in turquoise in the track diagram in their position in the genome, segments that are more similar to C. jejuni are shown in red, and equal segments in are shown in yellow. (b) Condensed representations of the Smash++ segments according to their similarity to C. coli and C. jejuni in absolute numbers and in per cent.
Fig. 4.
Fig. 4.
Functional subsystems identified in 141 genes of the hybrid strain meC0280 originating from C. jejuni . This pie chart illustrates the functional subsystems of the 141 genes of the hybrid strain meC0280, which presumably originated from C. jejuni . In particular, the two categories motility and chemotaxis as well as membrane transport dominate. Isolate meC0280 was chosen as an example because with 141 genes of putative C. jejuni origin compared to the other two strains, it contains the most genes of putative C. jejuni origin.
Fig. 5.
Fig. 5.
Venn diagram to illustrate the common proportions of genes putatively derived from C. jejuni and taken up by the hybrid strains. The three hybrid strains have 141, 136 or 124 genes integrated into their genome, which are presumably derived from C. jejuni . These genes are relatively different between the individual hybrid isolates although individual functional groups are clearly overrepresented. Just 23 genes originating from C. jejuni are found in all three hybrid genomes.
Fig. 6.
Fig. 6.
Methylation motifs in the three hybrid strains. Sequence logos of four methylation motifs detected in the hybrid isolate meC0280. The two motifs in A1 and in A2 as well as B1 and B2 are partner motifs that are methylated at both strands. All motifs are recognized by N-6 adenine-specific methyltransferases. The height of each stack indicates the degree of conservation (bits). The height of the letters represents the relative frequency of the base. The asterisk under a particular letter indicates the modified/methylated base.

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