Functional analysis of unique class II insertion sequence IS1071.

Masahiro Sota; Hirokazu Yano; Yuji Nagata; Yoshiyuki Ohtsubo; Hiroyuki Genka; Hisashi Anbutsu; Haruhiko Kawasaki; Masataka Tsuda

doi:10.1128/aem.72.1.291-297.2006

PubMed · 16391056

Functional analysis of unique class II insertion sequence IS1071.

Masahiro Sota·Hirokazu Yano·Yuji Nagata·Yoshiyuki Ohtsubo·Hiroyuki Genka·Hisashi Anbutsu·Haruhiko Kawasaki·Masataka Tsuda

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Abstract

Various xenobiotic-degrading genes on many catabolic plasmids are often flanked by two copies of an insertion sequence, IS1071. This 3.2-kb IS element has long (110-bp) terminal inverted repeats (IRs) and a transposase gene that are phylogenetically related to those of the class II transposons. However, the transposition mechanism of IS1071 has remained unclear. Our study revealed that IS1071 was only able to transpose at high frequencies in two environmental beta-proteobacterial strains, Comamonas testosteroni and Delftia acidovorans, and not in any of the bacteria examined which belong to the alpha- and gamma-proteobacteria. IS1071 was found to have the functional features of the class II transposons in that (i) the final product of the IS1071 transposition was a cointegrate of its donor and target DNA molecules connected by two directly repeated copies of IS1071, one at each junction; (ii) a 5-bp duplication of the target sequence was observed at the insertion site; and (iii) a tnpA mutation of IS1071 was efficiently complemented by supplying the wild-type tnpA gene in trans. Deletion analysis of the IS1071 IR sequences indicated that nearly the entire region of the IRs was required for its transposition, suggesting that the interaction between the transposase and IRs of IS1071 might be different from that of the other well-characterized class II transposons.

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Base Sequence Betaproteobacteria DNA Transposable Elements Molecular Sequence Data Plasmids Proteobacteria Sequence Analysis, DNA Terminal Repeat Sequences Transposases Journal Article Research Support, Non-U.S. Gov't

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Masahiro Sota, Hirokazu Yano, Yuji Nagata, Yoshiyuki Ohtsubo, Hiroyuki Genka, Hisashi Anbutsu, Haruhiko Kawasaki, Masataka Tsuda. 2006. Functional analysis of unique class II insertion sequence IS1071.. https://doi.org/10.1128/aem.72.1.291-297.2006

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