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Yeast Skn7p functions in a eukaryotic two-component regulatory pathway
J L Brown1, H Bussey, R C Stewart
1Department of Biology, McGill University, Montreal, Quebec, Canada.
The EMBO Journal
|November 1, 1994
Summary
Yeast Skn7p
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- The yeast SKN7 gene product, Skn7p, possesses a conserved receiver motif.
- This motif is homologous to bacterial response regulators involved in signal transduction.
- Skn7p's function in eukaryotes was previously uncharacterized.
Purpose of the Study:
- To investigate the role of the conserved receiver motif in Skn7p function.
- To determine the regulatory mechanism of Skn7p activity.
- To elucidate the relationship between Skn7p and the PKC1-MAP kinase pathway.
Main Methods:
- Site-directed mutagenesis of Skn7p at the conserved aspartate residue (D427).
- In vivo functional assays to assess Skn7p activity.
- Epistasis experiments to determine pathway interactions.
Main Results:
- Skn7p D427N mutation diminished activity, while D427E mutation enhanced activity, supporting a phosphorylation-dependent regulatory mechanism.
- Skn7p suppressed growth defects of a pkc1 delta mutation.
- Epistasis data indicated Skn7p functions in parallel to, not downstream of, the PKC1-MAP kinase pathway.
Conclusions:
- Experimental evidence supports the functional role of a receiver motif in regulating Skn7p activity in yeast.
- Skn7p likely functions as a transcription factor in a two-component signal transduction pathway.
- This pathway acts in parallel with the PKC1 cascade to regulate cell surface growth.