IHF is the limiting host factor in transposition of Pseudomonas putida transposon Tn4652 in stationary phase.
Transpositional activity of mobile elements is not constant. Conditional regulation of host factors involved in transposition may severely change the activity of mobile elements. We have demonstrated previously that transposition of Tn4652 in Pseudomonas putida is a stationary phase-specific event, which requires functional sigma S (Ilves et al., 2001, J Bacteriol 183: 5445-5448). We hypothesized that integration host factor (IHF), the concentration of which is increased in starving P. putida, might contribute to the transposition of Tn4652 as well. Here, we demonstrate that transposition of Tn4652 in stationary phase P. putida is essentially limited by the amount of IHF. No transposition of Tn4652 occurs in a P. putida ihfA-defective strain. Moreover, overexpression of IHF results in significant enhancement of transposition compared with the wild-type strain. This indicates that the amount of IHF is a bottleneck in Tn4652 transposition. Gel mobility shift and DNase I footprinting studies revealed that IHF is necessary for the binding of transposase to both transposon ends. In vitro, transposase can bind to inverted repeats of transposon only after the binding of IHF. The results obtained in this study indicate that, besides sigma S, IHF is another host factor that is implicated in the elevation of transposition in stationary phase.
Ilves
Heili
H
Estonian Biocentre and Institute of Molecular and Cell Biology, Tartu University, 23 Riia Street, 51010 Tartu, Estonia.
Hõrak
Rita
R
Teras
Riho
R
Kivisaar
Maia
M
D018983:DNA Footprinting; D004251:DNA Transposable Elements; D004591:Electrophoresis, Polyacrylamide Gel; D005798:Genes, Bacterial; D034982:Integration Host Factors / Q000378:metabolism*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D016958:Pseudomonas putida / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D014158:Transcription, Genetic; D019895:Transposases / Q000378:metabolism
D004251:DNA Transposable Elements; D034982:Integration Host Factors; C440503:integration host factor, Pseudomonas; D019895:Transposases