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Imaging G-protein Coupled Receptor GPCR-mediated Signaling Events that Control Chemotaxis of Dictyostelium Discoideum
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Phosphorylated Gβ is a directional cue during yeast gradient tracking.

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Yeast cells sense pheromone gradients to direct growth. Phosphorylated Gβ subunit dynamics, tracked by a novel biosensor, are crucial for this gradient sensing and mating efficiency.

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Area of Science:

  • Cell biology
  • Biochemistry
  • Genetics

Background:

  • Budding yeast cells use pheromone gradients to find mating partners.
  • A gradient-sensing model explains how yeast cells switch polarity sites.
  • Phosphorylation of the Gβ subunit is hypothesized to be key in this process.

Purpose of the Study:

  • To test predictions of a gradient-sensing model using a novel biosensor.
  • To investigate the spatiotemporal dynamics of phosphorylated Gβ during mating.
  • To determine the role of Gβ phosphorylation in yeast chemotropism.

Main Methods:

  • Development of a biosensor for phosphorylated Gβ.
  • Monitoring biosensor and reporter dynamics in mating yeast cells.
  • Analysis of cells expressing a nonphosphorylatable Gβ mutant.

Main Results:

  • The biosensor colocalized with Gβ and receptors, tracking from the default to the chemotropic site.
  • Phosphorylated Gβ concentrated on the leading side of Gβ/receptor peaks.
  • Nonphosphorylatable Gβ mutants showed impaired gradient tracking and mating.

Conclusions:

  • Gradient-regulated Gβ phosphorylation is essential for yeast gradient sensing.
  • Phosphorylated Gβ dynamics correlate with directional growth and mating partner orientation.
  • Gβ phosphorylation plays a critical role in mating efficiency.