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Related Experiment Videos

Interaction Dynamics Determine Signaling and Output Pathway Responses.

Klement Stojanovski1, Tony Ferrar2, Hannah Benisty2

  • 1Cell Signaling Research Group, Departament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, 08003 Barcelona, Spain.

Cell Reports
|April 6, 2017
PubMed
Summary

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Altering the association and dissociation rates of the Sln1-Ypd1 complex in the yeast high-osmolarity glycerol (HOG) pathway impacts signaling. Specific kinetic changes affect HOG pathway activation, basal signaling, and osmoadaptation.

Area of Science:

  • Cellular signaling
  • Biochemistry
  • Yeast genetics

Background:

  • Understanding signaling pathway dynamics is crucial for identifying therapeutic targets.
  • The yeast high-osmolarity glycerol (HOG)-mitogen-activated protein kinase (MAPK) pathway is a key osmosensor.
  • Kinetic rate constants play a significant role in biological processes.

Purpose of the Study:

  • To investigate the impact of kinetic rate constants (association, k_on; dissociation, k_off) of the Sln1-Ypd1 complex on HOG pathway signaling output.
  • To determine how manipulating these kinetic parameters affects osmosensing and adaptation in yeast.

Main Methods:

  • Created mutant pairs of the Sln1-Ypd1 complex interface to alter kinetic rate constants (k_on, k_off) while maintaining similar overall complex affinity (K_d).
Keywords:
HOG-MAPK pathwaykinetic perturbationsosmostress responsephosphorelay

Related Experiment Videos

  • Assessed HOG pathway activation thresholds, basal signaling, gene expression, and osmoadaptation in yeast cells expressing these mutants.
  • Utilized yeast genetics and molecular biology techniques to perturb and analyze the HOG-MAPK pathway.

    • Main Results:

    • Mutants with moderately increased k_on and k_off exhibited a lower threshold for HOG pathway activation compared to wild-type.
    • Mutants with higher k_on and k_off rates showed increased basal signaling and gene expression.
    • These higher kinetic rate mutants displayed impaired osmoadaptation, indicating a disruption in cellular response to osmotic stress.

    Conclusions:

    • The association (k_on) and dissociation (k_off) rates of the Sln1 osmosensor components are critical determinants of proper signaling dynamics.
    • Kinetic parameters, not just overall affinity, significantly influence the fidelity and effectiveness of cellular osmoadaptation responses.
    • Modulating specific kinetic rates offers a potential strategy for fine-tuning signaling pathways for therapeutic interventions.