Related Experiment Video

Updated: Jan 16, 2026

Characterize Disease-related Mutants of RAF Family Kinases by Using a Set of Practical and Feasible Methods
07:49

Characterize Disease-related Mutants of RAF Family Kinases by Using a Set of Practical and Feasible Methods

Published on: July 17, 2019

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Divergent Morphologies and Common Signaling Features of Active and Inactive Oncogenic RHOA Mutants in Yeast

Chenwei Wang1, Shinsuke Ohnuki1, Anna Savchenko1,2

  • 1Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa 277-8562, Japan.

Cells
|September 26, 2025
PubMed

Insights

Oncogenic RHOA mutations were studied in yeast, revealing distinct cellular effects based on mutation type. Morphology-based analysis effectively distinguished between gain-of-function and loss-of-function RHOA variants.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Cancer Research

Background:

  • Recurrent mutations in RHOA, a Rho family GTPase, are found in various cancers.
  • The cellular consequences of these RHOA mutations are not well understood.

Purpose of the Study:

  • To investigate the cellular effects of oncogenic RHOA variants using a yeast model.
  • To characterize the functional divergence of RHOA mutants.

Main Methods:

  • Expressed oncogenic RHOA variants in Saccharomyces cerevisiae, replacing the essential homolog RHO1.
  • Utilized high-dimensional image analysis to quantify 501 morphological parameters.
  • Applied principal component and linear discriminant analysis to discern phenotypic profiles.

Main Results:

  • Four RHOA variants (R5Q, G17V, C16R, A161P) were tested; E40Q did not complement RHO1 deletion.
  • All viable mutants conferred myriocin resistance, indicating membrane stress response activation.
  • Morphological profiling separated gain-of-function (C16R, A161P) from loss-of-function (R5Q, G17V) mutants.

Conclusions:

  • Yeast models can systematically dissect RHOA mutant functions.
  • Morphology-based approaches are valuable for characterizing context-dependent oncogenesis mechanisms.
  • Functional divergence of RHOA mutants was demonstrated through distinct phenotypic profiles.
Keywords:
CalMorphRHO1RHOAcanceryeast morphology

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