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Related Experiment Videos

Translational activation by an NtrC enhancer-binding protein

P J Cullen1, W C Bowman, D F Hartnett

  • 1Department of Biology, Washington University, St. Louis, MO 63130, USA.

Journal of Molecular Biology
|June 20, 1998
PubMed
Summary

The Rhodobacter capsulatus NtrC protein uniquely controls NifR3 production by stimulating translation, not transcription. This bacterial enhancer-binding protein

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Area of Science:

  • Bacterial gene regulation
  • Molecular biology
  • Protein-DNA interactions

Background:

  • The nifR3-ntrB-ntrC operon in Rhodobacter capsulatus encodes nitrogen-sensing regulators NtrB and NtrC, and the conserved NifR3 protein.
  • NtrC is a bacterial enhancer-binding protein known to activate transcription of several genes.
  • The function of NifR3 and its regulation by NtrC were previously unclear.

Purpose of the Study:

  • To investigate the mechanism by which NtrC regulates the production of NifR3 in Rhodobacter capsulatus.
  • To determine if NtrC controls NifR3 production at the transcriptional or translational level.
  • To elucidate the role of NtrC binding sites at the nifR3 promoter.

Main Methods:

  • LacZ reporter gene assays to analyze operon expression.

Related Experiment Videos

  • Immunological quantitation of NifR3 and NtrC protein levels.
  • In vitro binding assays with purified NtrC protein and the nifR3 promoter DNA.
  • Deletion analysis of NtrC binding sites in vivo.

    • Main Results:

    • NtrC regulates NifR3 production post-transcriptionally, impacting translation.
    • NtrC binds cooperatively to two sets of sites at the nifR3 promoter (+1 and -81).
    • Upstream binding sites are crucial for NtrC-dependent NifR3 production but not for transcription initiation.

    Conclusions:

    • NtrC stimulates NifR3 translation while bound to the nifR3 promoter DNA.
    • This represents a unique mechanism of translational control by an enhancer-binding protein.
    • NtrC's mode of action at the nifR3 promoter differs from its transcriptional activation of other genes.