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Related Experiment Videos

The yeast two-hybrid system detects interactions between Bacillus subtilis sigmaB regulators

U Voelker1, A Voelker, W G Haldenwang

  • 1Institute für Mikrobiologie und Molecularbiologié, Einst-Moritz-Arndt-Universität, Greifswald, Germany.

Journal of Bacteriology
|December 1, 1996
PubMed
Summary

The Bacillus subtilis general stress response regulator, sigmaB, is controlled by seven proteins. Yeast two-hybrid assays revealed new interactions among these regulatory proteins, expanding our understanding of sigmaB regulation.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • SigmaB is a key regulator of the general stress response in Bacillus subtilis.
  • Its activity is controlled by a complex regulatory cascade involving seven co-transcribed proteins (RsbR, RsbS, RsbT, RsbU, RsbV, RsbR, and RsbX).
  • Previous biochemical methods identified interactions between RsbW, RsbV, and sigmaB but failed to map other protein-protein interactions within the regulatory complex.

Purpose of the Study:

  • To investigate novel protein-protein interactions among the sigmaB regulatory cascade components.
  • To elucidate the interaction network governing sigmaB activation using a sensitive yeast two-hybrid system.

Main Methods:

  • Utilized the yeast two-hybrid system to screen for interactions between pairs of sigmaB regulatory proteins.

Related Experiment Videos

  • Tested interactions of RsbR, RsbS, RsbT, RsbU, RsbV, RsbR, and RsbX with each other and with sigmaB.

    • Main Results:

    • The yeast reporter system confirmed interactions between RsbS and RsbR or RsbT.
    • Interactions were also detected between RsbR and RsbT, and between RsbV and RsbU or RsbR.
    • Evidence for RsbR dimer formation (RsbR2) and RsbR2 dimer formation was observed.
    • RsbX did not interact with any other tested component of the sigB operon.

    Conclusions:

    • The yeast two-hybrid system successfully identified previously undetected interactions within the sigmaB regulatory network.
    • These findings provide a more comprehensive map of the protein interactions governing sigmaB activation in Bacillus subtilis.
    • The results highlight the complex regulatory mechanisms controlling the general stress response in bacteria.