Analysis of the N-terminal DNA binding domain of the IS30 transposase

Zita Nagy, Mónika Szabó, Michael Chandler, F. Olasz

Research output: Article

19 Citations (Scopus)

Abstract

IS30 is the founding member of a large family of widely spread bacterial insertion sequences with closely related transposases. The N-terminal end of the 1S30 transposase had been shown to retain sequence-specific DNA binding activity and to protect the IS30 terminal inverted repeats. Structural predictions revealed the presence of a helix-helix-turn-helix motif (H-HTH2) which, in the case of IS30, is preceded by an additional helix-turn-helix motif (HTH1). Analysis of deletion and point mutants in this region revealed that both motifs are important for IS30 transposition. IS30 exhibits two types of insertion specificity preferring either a 24 bp palindromic hot-spot (GOHS) or sequences resembling its ends [left and right terminal inverted repeat (IRL and IRR)]. Results are presented suggesting that the HTH1 region is required for GOHS targeting and interferes with the inverted repeat (IR) targeting. On the other hand, H-HTH2 appears to be required for both. The binding activities of the mutant proteins to the terminal IS30 IRs as measured by gel retardation correlated well with these results. Furthermore, close inspection of the H-HTH2 region revealed significant amino acid identity with a similar predicted secondary structure carried by the transcriptional regulator FixJ of Sinorhizobium meliloti and involved in FixJ binding to its target sequence. This suggests that FixJ and IS30 transposase share similar sequence-specific DNA binding mechanisms.

Original languageEnglish
Pages (from-to)478-488
Number of pages11
JournalMolecular Microbiology
Volume54
Issue number2
DOIs
Publication statusPublished - okt. 2004

Fingerprint

Transposases
Helix-Turn-Helix Motifs
Terminal Repeat Sequences
DNA
Sinorhizobium meliloti
Insertional Mutagenesis
Mutant Proteins
Gels
Amino Acids

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

Analysis of the N-terminal DNA binding domain of the IS30 transposase. / Nagy, Zita; Szabó, Mónika; Chandler, Michael; Olasz, F.

In: Molecular Microbiology, Vol. 54, No. 2, 10.2004, p. 478-488.

Research output: Article

Nagy, Zita ; Szabó, Mónika ; Chandler, Michael ; Olasz, F. / Analysis of the N-terminal DNA binding domain of the IS30 transposase. In: Molecular Microbiology. 2004 ; Vol. 54, No. 2. pp. 478-488.
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