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bioRxiv · 10.64898/2026.06.24.734251

Epigenomes in thermophilic microbial communities and their impact on the interaction across prokaryotes and mobilomes

Hiraoka, S.·Shimamura, S.·Usui, K.·Zhang, Y.·Sumida, T.·Tsukamoto, Y.·Kawai, S.·Nishihara, A.
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Abstract

DNA chemical modifications, including methylation, are widespread in prokaryotes and mobilomes, including viruses, plasmids, and other extrachromosomal DNAs, and play important roles in their ecology and interactions. However, current knowledge of these modification systems and their association with interactions between hosts and mobilomes across communities, including those in extreme environments, is severely limited. Here, using single-molecule real-time sequencing and single-cell genome sequencing technologies, we conducted a culture-independent metaepigenomic analysis of microbial communities in hot spring biofilms. A total of 248, 332, and 465 genomes were constructed from diverse prokaryotes, viruses, and extrachromosomal circular DNAs, respectively, from 10 biofilm samples collected from 3 hot spring sites. In total, 1106 candidate methylated motifs and 3280 genes associated with the restriction-modification (RM) system, including DNA methyltransferases (MTases), were identified. In contrast to the varied methylated motifs, the nucleotide-level modification ratios were consistent with those of a common Escherichia coli genome, and an environment-dependent epigenomic preference attributed to the lack of C5-methylcytosine was observed, as supported by direct measurements of modified bases by liquid chromatography-tandem mass spectrometry. A systematic survey revealed various defense systems in the genome, and almost half of the MTase genes were estimated to be genetically involved in defense mechanisms against extracellular DNA, such as RM systems. The mobilomes and their predicted hosts shared epigenomic patterns within each interactive subnetwork, suggesting that mobilome DNA was modified by host MTase during the current infection or transfection, rather than serving as historical records. Our findings highlight that DNA modification shapes multiple ecological and evolutionary strategies in interactions between prokaryotes and mobilomes, and that epigenomes serve as a potential signature for accurate prediction of current host-phage interactions.

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Hiraoka, S., Shimamura, S., Usui, K., Zhang, Y., Sumida, T., Tsukamoto, Y., Kawai, S., Nishihara, A.. 2026-06-25. Epigenomes in thermophilic microbial communities and their impact on the interaction across prokaryotes and mobilomes. https://doi.org/10.64898/2026.06.24.734251

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