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Conjugative transposition of the vancomycin resistance carrying Tn1549: enzymatic requirements and target site

Lotte Lambertsen1, Anna Rubio-Cosials1, Kiran Raosaheb Patil1

  • 1European Molecular Biology Laboratory (EMBL), Structural and Computational Biology Unit, Meyerhofstrasse 1, D-69117 Heidelberg, Germany.

Molecular Microbiology
|December 23, 2017
PubMed

Insights

The conjugative transposon Tn1549 can move between bacteria, facilitating antibiotic resistance transfer. Its movement relies on specific proteins and DNA sequences, enabling capture and transfer of host DNA.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Vancomycin resistance is a growing global health concern.
  • The conjugative transposon Tn1549 is often responsible for vancomycin resistance.
  • The mobility of Tn1549 between bacterial species is not fully understood.

Purpose of the Study:

  • To investigate the mobility of the conjugative transposon Tn1549.
  • To elucidate the molecular mechanisms underlying Tn1549 transposition.
  • To understand how Tn1549 facilitates antibiotic resistance transfer.

Main Methods:

  • Development of an in vivo assay system in E. coli.
  • Analysis of the roles of Tn1549-encoded INT and XIS proteins.
  • Investigation of DNA sequences flanking the transposon ends.

Main Results:

  • Mini-Tn1549 demonstrated transposition in E. coli, a Gram-negative host.
  • The INT enzyme and its catalytic tyrosine Y380 are essential for transposition.
  • The XIS protein is crucial for efficient and accurate transposition.
  • Specific DNA sequences flanking the left transposon end are critical for excision.
  • Mutations in flanking DNA can be compensated by changes at the right transposon end.
  • Tn1549 can capture and transfer flanking genomic DNA, especially when mobility is altered.
  • Integration target sites were identified as TTTT-N6-AAAA.

Conclusions:

  • Tn1549 exhibits conjugative transposition capabilities beyond its natural host.
  • Molecular insights into Tn1549 transposition mechanisms were provided.
  • The adaptability of Tn1549 for efficient antibiotic resistance transfer was highlighted.

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