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In vitro interactions of integration host factor with the ompF promoter-regulatory region of Escherichia coli

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Summary

Previous work has shown that integration host factor (IHF) mutants have increased expression and altered osmoregulation of OmpF, a major Escherichia coli outer membrane protein. By in vitro analysis the possibility was investigated that IHF interacts directly with the ompF promoter region. Gel retardation assays and DNase I protection experiments showed that IHF binds to two sites in the ompF promoter region centered at positions −180 and −60 relative to the start of transcription. Gel electrophoresis studies with circularly permuted ompF promoter fragments indicated that IHF binding strongly increased a small intrinsic bend in the ompF promoter region. The addition of IHF to a purified in vitro transcription system strongly and specifically inhibited ompF transcription. This inhibition was reversed by increasing the concentration of OmpR, a positive activator required for ompF expression, suggesting that IHF may inhibit ompF transcription by altering how OmpR interacts with the ompF promoter.

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Authors and Affiliations

  1. Department of Biochemistry and Cell Biology, State University of New York, 11794-5215, Stony Brook, NY, USA

    Natarajan Ramani, Lin Huang & Martin Freundlich

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  1. Natarajan Ramani
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  2. Lin Huang
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  3. Martin Freundlich
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Communicated by W. Arber

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Ramani, N., Huang, L. & Freundlich, M. In vitro interactions of integration host factor with the ompF promoter-regulatory region of Escherichia coli . Molec. Gen. Genet. 231, 248–255 (1992). https://doi.org/10.1007/BF00279798

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  • DOI: https://doi.org/10.1007/BF00279798

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