<jats:p>Transposition activity in bacteria is generally maintained at a low level. The activity of mobile DNA elements can be controlled by bacterially encoded global regulators. Regulation of transposition of Tn<jats:italic>4652</jats:italic>in<jats:italic>Pseudomonas putida</jats:italic>is one such example. Activation of transposition of Tn<jats:italic>4652</jats:italic>in starving bacteria requires the stationary-phase sigma factor RpoS and integration host factor (IHF). IHF plays a dual role in Tn<jats:italic>4652</jats:italic>translocation by activating transcription of the transposase gene<jats:italic>tnpA</jats:italic>of the transposon and facilitating TnpA binding to the inverted repeats of the transposon. Our previous results have indicated that besides IHF some other<jats:italic>P. putida</jats:italic>-encoded global regulator(s) might bind to the ends of Tn<jats:italic>4652</jats:italic>and regulate transposition activity. In this study, employing a DNase I footprint assay we have identified a binding site of<jats:italic>P. putida</jats:italic>Fis (factor for inversion stimulation) centred 135 bp inside the left end of Tn<jats:italic>4652</jats:italic>. Our results of gel mobility shift and DNase I footprint studies revealed that Fis out-competes IHF from the left end of Tn<jats:italic>4652</jats:italic>, thereby abolishing the binding of TnpA. Thus, the results obtained in this study indicate that the transposition of Tn<jats:italic>4652</jats:italic>is regulated by the cellular amount of<jats:italic>P. putida</jats:italic>global regulators Fis and IHF.</jats:p>