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Related Experiment Videos

Intrinsic bends and integration host factor binding at F plasmid oriT.

M M Tsai1, Y H Fu, R C Deonier

  • 1Department of Biological Sciences, University of Southern California, Los Angeles 90089-1340.

Journal of Bacteriology
|August 1, 1990
PubMed
Summary

This study analyzed the F plasmid oriT DNA for DNA bending and integration host factor (IHF) binding. Results show intrinsic DNA curvature and specific IHF binding sites, suggesting a structural role in oriT function.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • The F plasmid's origin of transfer (oriT) is crucial for bacterial conjugation.
  • Understanding oriT DNA structure and protein interactions is key to deciphering DNA transfer mechanisms.

Purpose of the Study:

  • To investigate the intrinsic DNA bending within the F plasmid oriT region.
  • To identify and characterize the binding sites of integration host factor (IHF) on oriT DNA.

Main Methods:

  • Permutation assays were used to detect intrinsic DNA bends.
  • Gel retardation and DNase footprinting assays were employed to study IHF binding.

Main Results:

  • A 527-bp fragment of F plasmid oriT exhibited intrinsic bending, with curvature centered around base pair 240.

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  • Two primary bending components were localized near base pairs 170 and 260.
  • Integration host factor (IHF) bound to a major site (bp 165-195) and a minor site (bp 287-319) within the oriT sequence.

    • Conclusions:

    • The intrinsic curvature of oriT DNA and IHF binding sites likely contribute to the structural organization and function of the origin of transfer.
    • These structural features may play a significant role in regulating plasmid conjugation initiation.