Involvement of two domains with helix-turn-helix and zinc finger motifs in the binding of IS1 transposase to terminal inverted repeats

Molecular Microbiology
Shinya OhtaE Ohtsubo

Abstract

The insertion element IS1 has two open reading frames (ORFs), insA and insB, and produces a transframe protein InsAB, known as IS1 transposase, by translational frameshifting. The transposase binds to terminal inverted repeats (IRL and IRR) to promote IS1 transposition. Unless frameshifting occurs, IS1 produces InsA protein, which also binds to IRs and therefore acts as an inhibitor of transposition, as well as a transcriptional repressor of the promoter in IRL. A helix-turn-helix (HTH) motif present in both transposase and InsA is thought to be involved in IR-specific DNA binding. A comparison of transposases encoded by IS1 family elements reveals that the N-terminal regions contain four conserved cysteine residues, which appear to constitute a C(2)C(2) zinc finger (ZF) motif. This motif is also thought to be involved in IR-specific DNA binding. In this study, we show that IS1 transposases with an amino acid substitution in the HTH or ZF motif lose the ability to promote transposition. We also show that transposases, as well as InsA proteins with the same substitution, lose the ability to repress the activity of the IRL promoter, and that purified InsA mutant proteins lose the ability to bind to the IRL-containing fragment. Fu...Continue Reading

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Citations

Mar 17, 2009·Research in Microbiology·Patricia SiguierMichael Chandler
Feb 7, 2014·FEMS Microbiology Reviews·Patricia SiguierMick Chandler
Jan 14, 2010·Critical Reviews in Biochemistry and Molecular Biology·Alison Burgess HickmanFred Dyda
Dec 14, 2011·Chembiochem : a European Journal of Chemical Biology·Biancamaria FarinaMaurizio Pellecchia
Jun 25, 2015·Microbiology Spectrum·Patricia SiguierMick Chandler

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