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Detection of Horizontal Gene Transfer Mediated by Natural Conjugative Plasmids in E. coli
Published on: March 24, 2023
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Recipient UvrD helicase is involved in single- to double-stranded DNA conversion during conjugative plasmid transfer
Minjia Shen1,2, Kelly Goldlust3, Sandra Daniel1
1Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198, Gif-sur-Yvette, France.
Nucleic Acids Research
|February 11, 2023
Summary
The UvrD protein is crucial for the successful transfer and establishment of antibiotic resistance plasmids between bacteria. Disabling UvrD hinders plasmid DNA conversion, impacting bacterial evolution and the spread of resistance.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Antibiotic resistance spread is a major societal challenge, often mediated by bacterial conjugation.
- Conjugation involves the transfer of single-stranded (ss) DNA, which must be converted to double-stranded (ds) DNA for plasmid establishment.
Purpose of the Study:
- To identify host-encoded factors influencing conjugative transfer frequency in Escherichia coli.
- To elucidate the role of the UvrD protein in plasmid establishment after conjugation.
Main Methods:
- Transposon insertion sequencing was used to screen for host factors affecting plasmid transfer.
- Analysis of UvrD mutants and live-cell microscopy were employed to study DNA conversion.
Main Results:
- Disruption of the recipient uvrD gene significantly reduced the frequency of conjugative plasmid acquisition.
- UvrD's ATPase activity, but not its dsDNA binding or RNA polymerase interaction, is essential for plasmid establishment.
- In uvrD mutants, transferred ssDNA often failed to convert to dsDNA.
Conclusions:
- UvrD plays a critical role in the establishment of horizontally acquired plasmid DNA, beyond its known function in genome integrity.
- UvrD is a key factor in the dynamics of plasmid transfer and the evolution of bacterial genome diversity.
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