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Comprehensive analysis of IncC plasmid conjugation identifies a crucial role for the transcriptional regulator AcaB.
- Source :
-
Nature microbiology [Nat Microbiol] 2020 Nov; Vol. 5 (11), pp. 1340-1348. Date of Electronic Publication: 2020 Aug 17. - Publication Year :
- 2020
-
Abstract
- The IncC family of broad-host-range plasmids enables the spread of antibiotic resistance genes among human enteric pathogens <superscript>1-3</superscript> . Although aspects of IncC plasmid conjugation have been well studied <superscript>4-9</superscript> , many roles of conjugation genes have been assigned based solely on sequence similarity. We applied hypersaturated transposon mutagenesis and transposon-directed insertion-site sequencing to determine the set of genes required for IncC conjugation. We identified 27 conjugation genes, comprising 19 that were previously identified (including two regulatory genes, acaDC) and eight not previously associated with conjugation. We show that one previously unknown gene, acaB, encodes a transcriptional regulator that has a crucial role in the regulation of IncC conjugation. AcaB binds upstream of the acaDC promoter to increase acaDC transcription; in turn, AcaDC activates the transcription of IncC conjugation genes. We solved the crystal structure of AcaB at 2.9-Šresolution and used this to guide functional analyses that reveal how AcaB binds to DNA. This improved understanding of IncC conjugation provides a basis for the development of new approaches to reduce the spread of these multi-drug-resistance plasmids.
- Subjects :
- Amino Acid Sequence
Crystallography, X-Ray
Drug Resistance, Multiple, Bacterial
Escherichia coli genetics
Escherichia coli Proteins chemistry
Escherichia coli Proteins genetics
Mutagenesis
Mutation
Promoter Regions, Genetic
Protein Structure, Secondary
Trans-Activators chemistry
Trans-Activators genetics
Transcription Factors chemistry
Transcription Factors genetics
Transcription, Genetic
Conjugation, Genetic genetics
Escherichia coli Proteins metabolism
Plasmids genetics
Trans-Activators metabolism
Transcription Factors metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 2058-5276
- Volume :
- 5
- Issue :
- 11
- Database :
- MEDLINE
- Journal :
- Nature microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 32807890
- Full Text :
- https://doi.org/10.1038/s41564-020-0775-0