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- Title
Conjugative transposition of the vancomycin resistance carrying Tn<italic>1549</italic>: enzymatic requirements and target site preferences.
- Authors
Lambertsen, Lotte; Rubio‐Cosials, Anna; Patil, Kiran Raosaheb; Barabas, Orsolya
- Abstract
Summary: Rapid spread of resistance to vancomycin has generated difficult to treat bacterial pathogens worldwide. Though vancomycin resistance is often conferred by the conjugative transposon Tn<italic>1549</italic>, it is yet unclear whether Tn<italic>1549</italic> moves actively between bacteria. Here we demonstrate, through development of an <italic>in vivo</italic> assay system, that a mini‐Tn<italic>1549</italic> can transpose in <italic>E. coli</italic> away from its natural Gram‐positive host. We find the transposon‐encoded INT enzyme and its catalytic tyrosine Y380 to be essential for transposition. A second Tn<italic>1549</italic> protein, XIS is important for efficient and accurate transposition. We further show that DNA flanking the left transposon end is critical for excision, with changes to nucleotides 7 and 9 impairing movement. These mutations could be partially compensated for by changing the final nucleotide of the right transposon end, implying concerted excision of the two ends. With changes in these essential DNA sequences, or without XIS, a large amount of flanking DNA transposes with Tn<italic>1549</italic>. This rescues mobility and allows the transposon to capture and transfer flanking genomic DNA. We further identify the transposon integration target sites as TTTT‐N6‐AAAA. Overall, our results provide molecular insights into conjugative transposition and the adaptability of Tn<italic>1549</italic> for efficient antibiotic resistance transfer.
- Subjects
VANCOMYCIN resistance; PATHOGENIC microorganisms; DNA; TRANSPOSASES; DRUG resistance in microorganisms
- Publication
Molecular Microbiology, 2018, Vol 107, Issue 5, p639
- ISSN
0950-382X
- Publication type
Article
- DOI
10.1111/mmi.13905