SGV1 encodes a CDC28/cdc2-related kinase required for a Gα subunit-mediated adaptive response to pheromone in S. cerevisiae

Irie, Kenji; Nomoto, Satoshi; Miyajima, Ikuko; Matsumoto, Kunihiro

doi:10.1016/0092-8674(91)90386-d

Cited by 78 publications

(63 citation statements)

References 43 publications

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“…Although thioacylation-defective Gpa1p binds to G ␤␥ complexes , the affinity of these subunit interactions has not been determined quantitatively. In mammalian systems, thioacylation of G s␣ increases its affinity for G ␤␥ subunits approximately fivefold (Irie et al, 1991). We found that modest increases in the expression of Gpa1p eliminated constitutive activity of the pathway (our unpublished observations).…”

Section: Plasma Membrane-targeting Via Lipid Modifications and Proteimentioning

confidence: 83%

Dual Lipid Modification Motifs in G_αand G_γSubunits Are Required for Full Activity of the Pheromone Response Pathway inSaccharomyces cerevisiae

Manahan

Patnana

Blumer

et al. 2000

MBoC

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To establish the biological function of thioacylation (palmitoylation), we have studied the heterotrimeric guanine nucleotide-binding protein (G protein) subunits of the pheromone response pathway of Saccharomyces cerevisiae. The yeast G protein ␥ subunit (Ste18p) is unusual among G ␥ subunits because it is farnesylated at cysteine 107 and has the potential to be thioacylated at cysteine 106. Substitution of either cysteine results in a strong signaling defect. In this study, we found that Ste18p is thioacylated at cysteine 106, which depended on prenylation of cysteine 107. Ste18p was targeted to the plasma membrane even in the absence of prenylation or thioacylation. However, G protein activation released prenylation-or thioacylation-defective Ste18p into the cytoplasm. Hence, lipid modifications of the G ␥ subunit are dispensable for G protein activation by receptor, but they are required to maintain the plasma membrane association of G ␤␥ after receptor-stimulated release from G ␣ . The G protein ␣ subunit (Gpa1p) is tandemly modified at its N terminus with amide-and thioester-linked fatty acids. Here we show that Gpa1p was thioacylated in vivo with a mixture of radioactive myristate and palmitate. Mutation of the thioacylation site in Gpa1p resulted in yeast cells that displayed partial activation of the pathway in the absence of pheromone. Thus, dual lipidation motifs on Gpa1p and Ste18p are required for a fully functional pheromone response pathway. INTRODUCTIONLipid modifications anchor heterotrimeric guanine nucleotide-binding proteins (G proteins) to the inner leaflet of the plasma membrane. G protein ␣ subunits are fatty acylated with amide-linked myristate, thioester-linked palmitate, or both. G protein ␥ subunits are prenylated with either farnesyl or geranylgeranyl moieties through stable thioether linkages. Prenylation of ␥ subunits and myristoylation of ␣ subunits are essential for the function of G proteins. These modifications promote plasma membrane association and facilitate high-affinity protein-protein interactions (reviewed in Wedegaertner et al., 1995). The functional consequences of thioacylation are less well understood. Thioacylation does not appear to be a major determinant of membrane avidity, at least in the presence of G ␤␥ subunits, but may play a role in targeting G ␣ specifically to the plasma membrane (Dunphy et al., 1996;Morales et al., 1998;Fishburn et al., 1999;Huang et al., 1999). In vitro studies have demonstrated the importance of thioester-linked lipid in mediating protein-protein interactions of G ␣ subunits. Thioacylation increases the affinity of G s␣ for G ␤␥ approximately fivefold (Iiri et al., 1996) and negatively regulates the interaction between regulators of G protein signaling and G ␣ subunits (Tu et al., 1997). Thus, thioacylation may impact protein-protein interactions, as well as the subcellular distribution of modified proteins.We have investigated thioacylation of G proteins in the genetically tractable organism Saccharomyces cerevisiae. The pheromone res...

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Section: Plasma Membrane-targeting Via Lipid Modifications and Proteimentioning

confidence: 83%

Dual Lipid Modification Motifs in G_αand G_γSubunits Are Required for Full Activity of the Pheromone Response Pathway inSaccharomyces cerevisiae

Manahan

Patnana

Blumer

et al. 2000

MBoC

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“…All the strains used in this study (Table 3) were derived from a wild-type strain KA31 (Irie et al 1991). Strain LG1, in which the GAL1 promoter was inserted together with the LEU2 marker into the SnaBI site at 53 bp upstream of the initiation codon of COF1, was obtained by one-step gene replacement as follows.…”

Section: Yeast Strainsmentioning

confidence: 99%

Cooperation of two actin‐binding proteins, cofilin and Aip1, in Saccharomyces cerevisiae

Iida

Yahara

1999

Genes to Cells

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Background: Cofilin is a low-molecular weight actinmodulating protein, and is structurally and functionally conserved among eukaryotes. Cofilin is encoded by COF1 in Saccharomyces cerevisiae, and is essential for cell viability. Cofilin binds to and severs actin filaments in vitro, and also enhances their depolymerization. A partner protein that cooperates with cofilin in vivo has not been identified.

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“…A halo bioassay was used to measure the sensitivity of cells to a factor-induced growth arrest as described (Irie et al 1991).…”

Section: Bioassay For Response To a Factormentioning

confidence: 99%

Functional analyses of mammalian protein kinase C isozymes in budding yeast and mammalian fibroblasts

Nomoto¹,

Watanabe

Ninomiya‐Tsuji

et al. 1997

Genes to Cells

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SGV1 encodes a CDC28/cdc2-related kinase required for a Gα subunit-mediated adaptive response to pheromone in S. cerevisiae

Cited by 78 publications

References 43 publications

Dual Lipid Modification Motifs in G_αand G_γSubunits Are Required for Full Activity of the Pheromone Response Pathway inSaccharomyces cerevisiae

Dual Lipid Modification Motifs in G_αand G_γSubunits Are Required for Full Activity of the Pheromone Response Pathway inSaccharomyces cerevisiae

Cooperation of two actin‐binding proteins, cofilin and Aip1, in Saccharomyces cerevisiae

Functional analyses of mammalian protein kinase C isozymes in budding yeast and mammalian fibroblasts

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SGV1 encodes a CDC28/cdc2-related kinase required for a Gα subunit-mediated adaptive response to pheromone in S. cerevisiae

Cited by 78 publications

References 43 publications

Dual Lipid Modification Motifs in Gαand GγSubunits Are Required for Full Activity of the Pheromone Response Pathway inSaccharomyces cerevisiae

Dual Lipid Modification Motifs in Gαand GγSubunits Are Required for Full Activity of the Pheromone Response Pathway inSaccharomyces cerevisiae

Cooperation of two actin‐binding proteins, cofilin and Aip1, in Saccharomyces cerevisiae

Functional analyses of mammalian protein kinase C isozymes in budding yeast and mammalian fibroblasts

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Dual Lipid Modification Motifs in G_αand G_γSubunits Are Required for Full Activity of the Pheromone Response Pathway inSaccharomyces cerevisiae

Dual Lipid Modification Motifs in G_αand G_γSubunits Are Required for Full Activity of the Pheromone Response Pathway inSaccharomyces cerevisiae