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PMID:8167411

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Citation

Ziman, M, Preuss, D, Mulholland, J, O'Brien, JM, Botstein, D and Johnson, DI (1993) Subcellular localization of Cdc42p, a Saccharomyces cerevisiae GTP-binding protein involved in the control of cell polarity. Mol. Biol. Cell 4:1307-16

Abstract

The Saccharomyces cerevisiae Cdc42 protein, a member of the Ras superfamily of low-molecular-weight GTP-binding proteins, is involved in the control of cell polarity during the yeast cell cycle. This protein has a consensus sequence (CAAX) for geranylgeranyl modification and is likely to be associated, at least in part, with cell membranes. Using cell fractionation and immunolocalization techniques, we have investigated the subcellular localization of Cdc42p. Cdc42p was found in both soluble and particulate pools, and neither its abundance nor its distribution varied through the cell cycle. The particulate form of Cdc42p could be solubilized with detergents but not with NaCl or urea, suggesting that it is tightly associated with membranes. An increase in soluble Cdc42p was observed in a geranylgeranyltransferase mutant strain (cdc43-2ts) grown at the restrictive temperature. In addition, Cdc42p from a cdc42C188S mutant strain (that has an alteration at the prenylation consensus site) was almost exclusively in the soluble fraction, suggesting that membrane localization is dependent on geranylgeranyl modification at Cys-188. Immunofluorescence and immunoelectron microscopy experiments demonstrated that Cdc42p localizes to the plasma membrane in the vicinity of secretory vesicles that were found at the site of bud emergence, at the tips and sides of enlarging buds, and within mating projections (shmoo tips) in alpha-factor-arrested cells. These results indicate that Cdc42p is localized to the bud site early in the cell cycle and suggest that this localization is critical for the selection of the proper site for bud emergence and for polarized cell growth.

Links

PubMed PMC275766

Keywords

Amino Acid Sequence; Cell Cycle; Cell Polarity; Consensus Sequence; Fungal Proteins/analysis; Fungal Proteins/physiology; GTP-Binding Proteins/analysis; GTP-Binding Proteins/physiology; Molecular Sequence Data; Protein Processing, Post-Translational; Reproduction, Asexual; Saccharomyces cerevisiae/chemistry; Saccharomyces cerevisiae/physiology; Saccharomyces cerevisiae/ultrastructure; Subcellular Fractions/chemistry; cdc42 GTP-Binding Protein, Saccharomyces cerevisiae

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

YEAST:CDC42

located_in

GO:0043332: mating projection tip

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

YEAST:CDC42

located_in

GO:0005886: plasma membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

YEAST:CDC42

located_in

GO:0005934: cellular bud tip

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

YEAST:CDC42

part_of

GO:0000131: incipient cellular bud site

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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