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Nucleotide sequence analysis of IS256 from the Staphylococcus aureus gentamicin-tobramycin-kanamycin-resistance

M E Byrne1, D A Rouch, R A Skurray

  • 1Department of Microbiology, Monash University, Clayton, Victoria, Australia.

Gene
|September 30, 1989
PubMed

Insights

The composite transposon Tn4001, common in Australian Staphylococcus aureus, carries aminoglycoside resistance genes. Sequencing IS256 elements completes the Tn4001 sequence, aiding understanding of antibiotic resistance.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Aminoglycoside resistance in Staphylococcus aureus, specifically gentamicin, tobramycin, and kanamycin (GmTmKmR), is frequently mediated by the composite transposon Tn4001.
  • The Tn4001 transposon harbors the aacA-aphD gene, encoding a bifunctional enzyme responsible for aminoglycoside modification.

Purpose of the Study:

  • To complete the entire nucleotide sequence of the Tn4001 transposon.
  • To analyze the sequence and structural features of the IS256 insertion sequences flanking the aacA-aphD gene.

Main Methods:

  • DNA sequencing of the IS256 elements.
  • Bioinformatic analysis of the IS256 sequence, including identification of open reading frames and terminal inverted repeats.
  • Integration of IS256 sequences with the previously reported aacA-aphD gene sequence.

Main Results:

  • The IS256 insertion sequences (IS256L and IS256R) flanking the aacA-aphD gene are identical and 1324 bp in length.
  • IS256 exhibits characteristic features of insertion sequence elements, including 26-bp imperfect terminal inverted repeats.
  • A single open reading frame within IS256 encodes a 45.6 kDa protein.
  • The complete sequence of Tn4001, totaling 4566 bp, is now determined.

Conclusions:

  • The full nucleotide sequence of Tn4001 has been elucidated.
  • Understanding the IS256 elements provides insights into the structure and potential mobility of Tn4001.
  • This complete sequence data is crucial for further studies on aminoglycoside resistance mechanisms in Staphylococcus aureus.

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