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Journal of Bacteriology, February 2000, p. 589-598, Vol. 182, No. 3
Department of Genetics, Institute of
Molecular and Cell Biology, Estonian Biocentre and Tartu
University, 51010 Tartu, Estonia
Received 27 May 1999/Accepted 31 October 1999
We have previously shown that both ends of the Tn3
family transposon Tn4652 contain integration host factor
(IHF) binding sites and that IHF positively regulates expression of the
Tn4652 transposase gene tnpA in
Pseudomonas putida (R. Hõrak, and M. Kivisaar, J. Bacteriol. 180:2822-2829, 1998). Tn4652 can activate silent genes by creating fusion promoters during the transposition. The
promoters are created as fusions between the
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Transcription from Fusion Promoters Generated during
Transposition of Transposon Tn4652 Is Positively Affected by
Integration Host Factor in Pseudomonas putida
35 hexamer provided by
the terminal inverted repeats of Tn4652 and the 10
hexamers in the target DNA. Two fusion promoters, PRA1 and PLA1, that
contain sequences of the right and left termini of Tn4652,
respectively, were chosen for the study of mechanisms of transcription
activation. Gel mobility shift analysis using crude extracts from
P. putida cells allowed us to detect specific binding of
P. putida IHF to the ends of the transposon
Tn4652. We found that the rate of transcription from the
fusion promoter PRA1 is enhanced by IHF. Notably, the positive effect
of IHF on transcription from the promoter PRA1 appeared only when cells
of P. putida reached the stationary growth phase. We
speculate that the intracellular concentration of IHF might be critical
for the in vivo effect of IHF on transcription from the fusion
promoters in P. putida. In the case of PLA1, the mechanism
of transcription modulation by IHF is different than that observed for
PRA1. Our results demonstrate that transcription of neighboring genes
from outwardly directed promoters at the ends of a mobile DNA
element could be influenced by the same factors that control
transposition of the element.
*
Corresponding author. Mailing address: Department of
Genetics, Institute of Molecular and Cell Biology, Estonian Biocentre and Tartu University, 23 Riia St., 51010 Tartu, Estonia. Phone: 372-7-375015. Fax: 372-7-420286. E-mail: maiak{at}ebc.ee.
Journal of Bacteriology, February 2000, p. 589-598, Vol. 182, No. 3
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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