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Updated: Jan 11, 2026
Phage Phenomics: Physiological Approaches to Characterize Novel Viral Proteins
Published on: June 11, 2015
Virion content unpacked by long-read sequencing: stress-induced changes in transmitted staphylococcal mobilome due to
Tibor Botka1, Soňa Smetanová1,2, Adam Vinco1
1Department of Experimental Biology, Faculty of Science, Masaryk University, Brno 625 00, Czech Republic.
Abstract:
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.
Insights
This study reveals how mobile genetic elements spread in Staphylococcus bacteria. Lytic phage therapy does not increase the risk of transferring these elements, offering insights into antibiotic resistance.
Area of Science:
- Microbiology
- Genetics
- Molecular Biology
Background:
- Staphylococci virulence and antibiotic resistance are influenced by their mobilome.
- Mobile genetic elements spread via phage virions and phage satellites (PICIs).
- Analyzing DNA content at the single-virion level for transducing particles has been challenging.
Purpose of the Study:
- To analyze the DNA content of transducing particles at the single-virion level.
- To investigate the impact of helper phage and antimicrobials on packaged mobilome composition.
- To assess the risk of mobile genetic element transfer with lytic phage therapy.
Main Methods:
- Utilized Staphylococcus epidermidis as a model organism.
- Employed long-read nanopore sequencing for DNA analysis.
- Determined the sequence structure and ratio of packaged DNA in virions.
Main Results:
- Characterized the DNA content (phage, PICI, plasmid, bacterial) in normal and small-headed virions.
- Observed that DNA packaging ratios depend on the helper phage and antimicrobial induction.
- Found no significant increase in mobilome dissemination with lytic phage and ciprofloxacin compared to antibiotics alone.
Conclusions:
- Developed a novel approach for analyzing the transduced bacterial mobilome.
- Demonstrated that lytic phage-based therapeutic strategies do not elevate the risk of mobile genetic element transfer in vitro.
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