Recipient UvrD helicase is involved in single- to double-stranded DNA conversion during conjugative plasmid transfer.
Minjia ShenKelly GoldlustSandra DanielChristian LesterlinYoshiharu YamaichiPublished in: Nucleic acids research (2023)
Dissemination of antibiotic resistance, a current societal challenge, is often driven by horizontal gene transfer through bacterial conjugation. During conjugative plasmid transfer, single-stranded (ss) DNA is transferred from the donor to the recipient cell. Subsequently, a complete double-stranded (ds) plasmid molecule is generated and plasmid-encoded genes are expressed, allowing successful establishment of the transconjugant cell. Such dynamics of transmission can be modulated by host- or plasmid-encoded factors, either in the donor or in the recipient cell. We applied transposon insertion sequencing to identify host-encoded factors that affect conjugative transfer frequency in Escherichia coli. Disruption of the recipient uvrD gene decreased the acquisition frequency of conjugative plasmids belonging to different incompatibility groups. Results from various UvrD mutants suggested that dsDNA binding activity and interaction with RNA polymerase are dispensable, but ATPase activity is required for successful plasmid establishment of transconjugant cells. Live-cell microscopic imaging showed that the newly transferred ssDNA within a uvrD- recipient often failed to be converted to dsDNA. Our work suggested that in addition to its role in maintaining genome integrity, UvrD is also key for the establishment of horizontally acquired plasmid DNA that drives genome diversity and evolution.
Keyphrases
- escherichia coli
- crispr cas
- single cell
- genome wide
- cell therapy
- klebsiella pneumoniae
- circulating tumor
- binding protein
- cell free
- nucleic acid
- single molecule
- biofilm formation
- antibiotic resistance genes
- stem cells
- high resolution
- induced apoptosis
- gene expression
- signaling pathway
- dna methylation
- cell death
- multidrug resistant
- pseudomonas aeruginosa
- bone marrow
- wild type