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Assay for Adhesion and Agar Invasion in S. cerevisiae
Published on: November 8, 2006
Multiple proteins and phosphorylations regulate Saccharomyces cerevisiae Cdc24p localization
Karen C Cole1, Joy-El R Barbour, John F Midkiff
1Department of Microbiology and Molecular Genetics, University of Vermont, 202 Stafford Hall, 95 Carrigan Drive, VT 05405, United States.
FEBS Letters
|September 29, 2009
Summary
Proper targeting of Saccharomyces cerevisiae Cdc24p to growth sites is crucial. This study identifies key interacting proteins and phosphorylation sites involved in directing Cdc24p localization for cell growth.
Area of Science:
- Cell biology
- Molecular and developmental biology
Background:
- Targeting of the Cdc24p protein to specific cellular locations is vital for its function in Saccharomyces cerevisiae.
- Understanding the factors that regulate Cdc24p localization is key to deciphering its role in polarized growth.
Purpose of the Study:
- To investigate the roles of Cdc24p-interacting proteins in its localization.
- To determine the impact of specific Cdc24p phosphorylation sites on its targeting to polarized growth sites.
Main Methods:
- Localization assays of GFP-tagged Cdc24 proteins and fragments.
- Analysis in deletion mutants of known Cdc24p-interacting proteins (boi2Δ, ent2Δ, hua1Δ, tos2Δ skg6Δ).
- Site-directed mutagenesis of potential Cdc24p phosphorylation sites (Ser697, Thr704, Tyr200).
Main Results:
- Deletion mutants boi2Δ, ent2Δ, and hua1Δ exhibited Cdc24p localization defects.
- The tos2Δ skg6Δ double mutant showed aberrant pre-anaphase localization of Cdc24p to the mother-bud neck.
- Mutations at Ser697, Thr704, and Tyr200 mimicked the aberrant localization pattern, with S697A also causing in vivo phosphorylation defects.
Conclusions:
- Boi2p, Ent2p, Hua1p, and Tos2p are important for targeting Cdc24p to polarized growth sites.
- Cdc24p phosphorylation, particularly at Ser697, plays a role in its proper localization.
- These findings elucidate novel regulatory mechanisms for Cdc24p function in yeast cell polarity.
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