Analysis in vitro and in vivo of the transcriptional regulator CrgA of Neisseria meningitidis upon contact with target cells - Institut Pasteur Access content directly
Journal Articles Molecular Microbiology Year : 2004

Analysis in vitro and in vivo of the transcriptional regulator CrgA of Neisseria meningitidis upon contact with target cells

Ala-Eddine Deghmane
Dario Giorgini
  • Function : Author
Laure Maigre
  • Function : Author
Muhamed-Kheir Taha
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Abstract

Contact between CrgA, a LysR-like regulatory protein in Neisseria meningitidis, and DNA is involved in the repression of several bacterial genes upon contact with epithelial cells. We used a defined in vitro system containing crgA promoter, purified RNA polymerase (RNAP) and purified CrgA protein to demonstrate that CrgA was directly responsible for this transcriptional repression. Interaction between the C-terminal domain of CrgA and the RNAP led to the production of short abortive transcripts, suggesting that CrgA may act by preventing RNAP from clearing the promoter. We probed the regulation by CrgA of its own production by analysing CrgA-DNA contacts during cell-bacteria interaction by assaying in vivo protection against dimethyl sulphate (DMS) methylation. Comparison of DMS footprints in vitro and in vivo suggested that CrgA repressed transcription through specific base contacts, probably in the major groove of the DNA double helix, resulting in DNA looping. Upon contact with target cells, CrgA was released from the DNA, allowing transcription of the target gene to proceed to elongation and facilitating tight control of the expression of genes regulated by CrgA.

Domains

Bacteriology

Dates and versions

pasteur-03262247 , version 1 (16-06-2021)

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Cite

Ala-Eddine Deghmane, Dario Giorgini, Laure Maigre, Muhamed-Kheir Taha. Analysis in vitro and in vivo of the transcriptional regulator CrgA of Neisseria meningitidis upon contact with target cells. Molecular Microbiology, 2004, 53 (3), pp.917-927. ⟨10.1111/j.1365-2958.2004.04167.x⟩. ⟨pasteur-03262247⟩

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