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Functions for Cdc42p BEM adaptors in regulating a differentiation-type MAP kinase pathway.

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|January 16, 2020
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This study reveals unique roles for Rho GTPase adaptors Bem4p and Bem1p in regulating the filamentous growth mitogen-activated protein kinase (fMAPK) pathway in yeast. Findings highlight sequential adaptor functions in controlling cell polarity and MAPK signaling.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Ras homology (Rho) GTPases, like Cdc42p in yeast, are crucial regulators of cell polarity and signal transduction.
  • Cdc42p influences multiple mitogen-activated protein kinase (MAPK) pathways, including mating, filamentous growth (fMAPK), and high osmolarity glycerol (HOG).
  • The precise mechanisms by which Cdc42p regulates the fMAPK pathway remain incompletely understood.

Purpose of the Study:

  • To elucidate the specific roles of Cdc42p adaptors, Bem4p and Bem1p, in regulating the yeast fMAPK pathway.
  • To compare the activation kinetics of different Cdc42p-dependent MAPK pathways.
  • To identify key interactions and domains within Bem4p and Bem1p essential for fMAPK signaling.

Main Methods:

  • Comparative analysis of MAPK pathway activation profiles in a filamentous yeast strain.
  • Site-directed mutagenesis to assess the function of specific Cdc42p and Bem4p residues.
  • Investigation of Bem1p mutants defective in plasma membrane recruitment, intramolecular interactions, and GEF interaction.
  • Genetic suppression tests to determine the order of Bem4p and Bem1p function.

Main Results:

  • The fMAPK pathway exhibits slower activation kinetics compared to mating and HOG pathways.
  • Cdc42pE100A, defective in fMAPK signaling, shows impaired interaction with the adaptor Bem4p.
  • Specific Bem4p residues are critical for Cdc42p interaction and fMAPK pathway signaling.
  • Bem1p's function in the fMAPK pathway depends on its ability to recruit Ste20p, its intramolecular interactions, and its interaction with the GEF Cdc24p.
  • Bem4p and Bem1p function sequentially in regulating the fMAPK pathway.

Conclusions:

  • Rho GTPase adaptors exhibit unique and sequential functions in regulating distinct MAPK pathways.
  • Bem4p and Bem1p play critical, ordered roles in the Cdc42p-dependent fMAPK signaling cascade.
  • Understanding these adaptor functions provides insight into the complex regulation of cell polarity and morphogenesis.