Virion content unpacked by long-read sequencing: Stress-induced changes in transmitted staphylococcal mobilome due to phage-satellite interactions

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Authors

BOTKA Tibor SMETANOVÁ Soňa VINCO Adam KUČEROVÁ Eliška ROVŇÁKOVÁ Kristína SIVÁKOVÁ Alena MAŠLAŇOVÁ Ivana PANTŮČEK Roman

Year of publication 2025
Type Article in Periodical
Magazine / Source NUCLEIC ACIDS RESEARCH
MU Faculty or unit

Faculty of Science

Citation
web https://doi.org/10.1093/nar/gkaf1165
Doi https://doi.org/10.1093/nar/gkaf1165
Keywords Mobile Genetic Elements; Nanopore Sequencing; Horizontal Gene Transfer; Phage Therapy; DNA packaging; Bacteriophage Satellites
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Description The evolution of the virulence and antibiotic resistance of staphylococci, important opportunistic pathogens, is strongly determined by their mobilome, which can spread by phage virions or small-headed particles resulting from the hijacking of helper phage machinery by phage satellites named phage-inducible chromosomal islands (PICIs). Despite known mechanisms of the formation of transducing particles, it has not yet been possible to analyze their DNA content at the single-virion level. Using the Staphylococcus epidermidis model and long-read nanopore sequencing, we determined the sequence structure and ratio of phage and PICI genophores, plasmid, and bacterial DNA packaged in normal and small-headed virions. It was shown that the ratios vary mainly depending on the helper phage and the antimicrobial used for induction. When the effect of a strictly lytic phage and its combination with ciprofloxacin on a packaged mobilome was analyzed, no significant increase in mobilome dissemination was observed compared to antibiotics alone. Here, we demonstrate a novel approach for the analysis of transduced bacterial mobilome and show in vitro that lytic phage-based therapeutic strategies do not increase the risk of mobile genetic element transfer.
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