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Yeasts are single-celled organisms, but unlike bacteria, they are eukaryotes (cells with a nucleus). Cell signaling in yeast is similar to signaling in other eukaryotic cells. A ligand, such as a protein or a small molecule released from a yeast cell, attaches to a receptor on the cell surface. The binding stimulates second-messenger kinases to activate or inactivate transcription factors that further regulate gene expression. Many of the yeast intracellular signaling cascades have similar...
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Manipulation and Analysis of Cell Cycle-Dependent Processes in Budding Yeast
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Published on: September 26, 2025

647

Probing Cdc42 Polarization Dynamics in Budding Yeast Using a Biosensor.

Satoshi Okada1, Mid Eum Lee2, Erfei Bi3

  • 1University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, United States; Kyushu University Graduate School of Medical Sciences, Fukuoka, Japan.

Methods in Enzymology
|March 25, 2017
PubMed
Summary
This summary is machine-generated.

Researchers developed a biosensor to track active Cdc42 (a GTPase) in yeast. This tool monitors Cdc42 activation in vivo, aiding the study of cell polarity and GTPase signaling.

Keywords:
CRIBCdc42FRAPGEFGFPGTPase

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Cdc42, a small GTPase, is crucial for cell polarity.
  • Dynamic spatiotemporal control of Cdc42 activity necessitates in vivo monitoring tools.
  • Understanding GTPase signaling pathways is vital in diverse biological contexts.

Purpose of the Study:

  • To construct and validate a biosensor for monitoring active Cdc42 in budding yeast.
  • To provide a tool for real-time visualization of Cdc42 activation dynamics.
  • To enable the study of Cdc42's role in cellular processes.

Main Methods:

  • Development of an affinity-based biosensor.
  • Fusion of a red fluorescent protein with a Cdc42- and Rac-interactive binding (CRIB) motif.
  • Application of the biosensor in budding yeast for in vivo monitoring.

Main Results:

  • A robust and simple biosensor for active, GTP-bound Cdc42 was successfully constructed.
  • The biosensor specifically binds to and reports on Cdc42 activation in vivo.
  • Demonstrated utility in monitoring Cdc42 dynamics in budding yeast.

Conclusions:

  • The developed biosensor is effective for studying Cdc42 activation in budding yeast.
  • This tool can be adapted for studying GTPase signaling in other cell types due to conserved CRIB motifs.
  • Facilitates research into cell polarity and signaling pathways involving Cdc42.