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Molecular Genetics and Genomics

, Volume 270, Issue 1, pp 94–102 | Cite as

H-NS and Lrp serve as positive modulators of traJ expression from the Escherichia coli plasmid pRK100

  • M. Starčič-Erjavec
  • J. P. M. van Putten
  • W. Gaastra
  • B. J. A. M. Jordi
  • M. Grabnar
  • D. Žgur-BertokEmail author
Original Paper

Abstract

Conjugative transfer of F-like plasmids is a tightly regulated process. The TraJ protein is the main positive activator of the tra operon which encodes products required for conjugative transfer of F-like plasmids. Nucleotide sequence analysis revealed potential Lrp and H-NS binding sites in the traJ regulatory region. Expression of a traJ-lacZ fusion in hns and lrp mutant strains showed that both are positive modulators of traJ expression. Competitive RT-PCR demonstrated that H-NS and Lrp exert their effect at the transcriptional level. Electrophoretic mobility-shift assays showed that H-NS and Lrp proteins bind to the traJ promoter. Conjugative transfer of pRK100 was decreased in hns but not in lrp mutant strains. Together, the results indicate H-NS and Lrp function as activators of traJ transcription.

Keywords

TraJ Lrp H-NS Conjugation F-like plasmids 

Notes

Acknowledgements

The authors thank Travis Tani for the Lrp protein, Rowena Matthews for plasmid pBE18 with the gltBDF promoter, and Sylvie Rimsky for the H-NS protein. The authors also thank Jerica Sabotič and Peter Mrak for help with performing the mating assays and Irena Kuhar for fruitful discussions and support. B.J.A.M. Jordi is a recipient of a fellowship of the Royal Netherlands Academy of Arts and Sciences

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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • M. Starčič-Erjavec
    • 1
  • J. P. M. van Putten
    • 2
  • W. Gaastra
    • 2
  • B. J. A. M. Jordi
    • 2
  • M. Grabnar
    • 1
  • D. Žgur-Bertok
    • 1
    Email author
  1. 1.Department of Biology, Biotechnical FacultyUniversity of LjubljanaLjubljanaSlovenia
  2. 2.Bacteriology Division, Department of Infectious Diseases and ImmunologyUtrecht UniversityUtrechtThe Netherlands

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