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Functional analysis of unique class II insertion sequence IS1071.

Masahiro Sota1, Hirokazu Yano, Yuji Nagata

  • 1Department of Biological Sciences, 222 Life Sciences North, University of Idaho, Moscow, ID 83844-3051, USA. sota@uidaho.edu

Applied and Environmental Microbiology
|January 5, 2006
PubMed
Summary

The insertion sequence IS1071, found near xenobiotic-degrading genes, can transpose in specific beta-proteobacteria. Its transposition mechanism shares features with class II transposons but may involve unique transposase-inverted repeat interactions.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Xenobiotic-degrading genes are often located on catabolic plasmids.
  • These genes are frequently flanked by the insertion sequence IS1071.
  • IS1071 shares characteristics with class II transposons, but its transposition mechanism is not fully understood.

Purpose of the Study:

  • To investigate the transposition mechanism of the insertion sequence IS1071.
  • To determine the host range and functional features of IS1071 transposition.
  • To explore the role of IS1071's terminal inverted repeats (IRs) in transposition.

Main Methods:

  • Transposition assays were performed in various bacterial strains (beta-, alpha-, and gamma-proteobacteria).
  • Analysis of transposition products, including cointegrates and target sequence duplication.

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  • Complementation experiments using a wild-type tnpA gene.
  • Deletion analysis of IS1071 IR sequences.

    • Main Results:

    • IS1071 transposed efficiently only in Comamonas testosteroni and Delftia acidovorans (beta-proteobacteria).
    • Transposition resulted in cointegrates and a 5-bp target sequence duplication, characteristic of class II transposons.
    • A mutation in the tnpA gene could be complemented in trans.
    • Nearly complete IR regions were essential for IS1071 transposition.

    Conclusions:

    • IS1071 functions as a class II transposon but exhibits a restricted host range within beta-proteobacteria.
    • The transposition mechanism of IS1071 may involve unique interactions between its transposase and IRs compared to other class II transposons.
    • Further research is needed to elucidate the specific molecular interactions governing IS1071 transposition.