Genome-Scale Analysis Reveals Sst2 as the Principal Regulator of Mating Pheromone Signaling in the Yeast Saccharomyces cerevisiae

ABSTRACT A common property of G protein-coupled receptors is that they become less responsive with prolonged stimulation. Regulators of G protein signaling (RGS proteins) are well known to accelerate G protein GTPase activity and do so by stabilizing the transition state conformation of the G protein α subunit. In the yeast Saccharomyces cerevisiae there are four RGS-homologous proteins (Sst2, Rgs2, Rax1, and Mdm1) and two Gα proteins (Gpa1 and Gpa2). We show that Sst2 is the only RGS protein that binds selectively to the transition state conformation of Gpa1. The other RGS proteins also bind Gpa1 and modulate pheromone signaling, but to a lesser extent and in a manner clearly distinct from Sst2. To identify other candidate pathway regulators, we compared pheromone responses in 4,349 gene deletion mutants representing nearly all nonessential genes in yeast. A number of mutants produced an increase (sst2, bar1, asc1, and ygl024w) or decrease (cla4) in pheromone sensitivity or resulted in pheromone-independent signaling (sst2, pbs2, gas1, and ygl024w). These findings suggest that Sst2 is the principal regulator of Gpa1-mediated signaling in vivo but that other proteins also contribute in distinct ways to pathway regulation.

[1]  P J Cullen,et al.  Defects in protein glycosylation cause SHO1-dependent activation of a STE12 signaling pathway in yeast. , 2000, Genetics.

[2]  R. W. Davis,et al.  A dominant truncation allele identifies a gene, STE20, that encodes a putative protein kinase necessary for mating in Saccharomyces cerevisiae. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[3]  Ronald W. Davis,et al.  Functional profiling of the Saccharomyces cerevisiae genome , 2002, Nature.

[4]  R. Aebersold,et al.  RGSZ1, a Gz-selective RGS Protein in Brain , 1998, The Journal of Biological Chemistry.

[5]  Peter Uetz,et al.  The yeast G protein alpha subunit Gpa1 transmits a signal through an RNA binding effector protein Scp160. , 2003, Molecular cell.

[6]  J. Benovic,et al.  G Protein-coupled Receptor Kinase 2/Gαq/11Interaction: A NOVEL SURFACE ON A REGULATOR OF G PROTEIN SIGNALING HOMOLOGY DOMAIN FOR BINDING Gα SUBUNITS , 2003 .

[7]  J. Heitman,et al.  The G protein-coupled receptor gpr1 is a nutrient sensor that regulates pseudohyphal differentiation in Saccharomyces cerevisiae. , 2000, Genetics.

[8]  H. Hamm,et al.  GTPase mechanism of Gproteins from the 1.7-Å crystal structure of transducin α - GDP AIF−4 , 1994, Nature.

[9]  R. Aebersold,et al.  Feedback Phosphorylation of an RGS Protein by MAP Kinase in Yeast* , 1999, The Journal of Biological Chemistry.

[10]  A. Hinnebusch,et al.  The WD protein Cpc2p is required for repression of Gcn4 protein activity in yeast in the absence of amino‐acid starvation , 1999, Molecular microbiology.

[11]  J. Moskow,et al.  Role of Cdc42p in Pheromone-Stimulated Signal Transduction in Saccharomyces cerevisiae , 2000, Molecular and Cellular Biology.

[12]  M. Whiteway,et al.  Theprotein kinase homologue Ste2Opisrequired tolink theyeastpheromone response G-protein flysubunits to downstream signalling components , 1992 .

[13]  A. Link,et al.  Yeast Asc1p and Mammalian RACK1 Are Functionally Orthologous Core 40S Ribosomal Proteins That Repress Gene Expression , 2004, Molecular and Cellular Biology.

[14]  Allen D. Roses,et al.  Genome-based pharmacogenetics and the pharmaceutical industry , 2002, Nature Reviews Drug Discovery.

[15]  A. Levey,et al.  RGS2 Binds Directly and Selectively to the M1 Muscarinic Acetylcholine Receptor Third Intracellular Loop to Modulate Gq/11α Signaling* , 2004, Journal of Biological Chemistry.

[16]  G. Sprague,,et al.  The roles of bud-site-selection proteins during haploid invasive growth in yeast. , 2002, Molecular biology of the cell.

[17]  R. Aebersold,et al.  RGSZ1, a G(z)-selective rgs protein in brain: Structure, membrane association, regulation by Gα(z) phosphorylation, and relationship to a G(z) gtpase-activating protein subfamily , 1998 .

[18]  P. Insel,et al.  RGS-PX1, a GAP for Gαs and Sorting Nexin in Vesicular Trafficking , 2001, Science.

[19]  David,et al.  RGS4 and GAIP are GTPase-activating proteins for G q (cid:97) and block activation of phospholipase C (cid:98) by (cid:103) -thio-GTP-G q (cid:97) , 1997 .

[20]  H. Hamm,et al.  The βγ Subunit of Heterotrimeric G Proteins Interacts with RACK1 and Two Other WD Repeat Proteins* , 2002, The Journal of Biological Chemistry.

[21]  D. Siderovski,et al.  D2 dopamine receptor activation of potassium channels is selectively decoupled by Gαi‐specific GoLoco motif peptides , 2005, Journal of neurochemistry.

[22]  M. Whiteway,et al.  Pheromone signalling and polarized morphogenesis in yeast. , 1997, Current opinion in genetics & development.

[23]  H. Dohlman,et al.  Analysis of RGS proteins in Saccharomyces cerevisiae. , 2002, Methods in enzymology.

[24]  H. Dohlman,et al.  RGS proteins: G protein-coupled receptors meet their match. , 2003, Assay and drug development technologies.

[25]  S. Sprang,et al.  Structure of RGS4 Bound to AlF4 −-Activated Giα1: Stabilization of the Transition State for GTP Hydrolysis , 1997, Cell.

[26]  Ronald W. Davis,et al.  Functional characterization of the S. cerevisiae genome by gene deletion and parallel analysis. , 1999, Science.

[27]  K. Blumer,et al.  Pak-Family Kinases Regulate Cell and Actin Polarization Throughout the Cell Cycle of Saccharomyces cerevisiae , 1999, Journal of Cell Biology.

[28]  J. Douwes,et al.  Loss of the Plasma Membrane-Bound Protein Gas1p inSaccharomyces cerevisiae Results in the Release of β1,3-Glucan into the Medium and Induces a Compensation Mechanism To Ensure Cell Wall Integrity , 1998, Journal of bacteriology.

[29]  A. Sands,et al.  Knockouts model the 100 best-selling drugs—will they model the next 100? , 2003, Nature Reviews Drug Discovery.

[30]  A. Gilman,et al.  Aluminum: a requirement for activation of the regulatory component of adenylate cyclase by fluoride. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[31]  J. Kurjan,et al.  The yeast SCG1 gene: A Gα-like protein implicated in the a- and α-factor response pathway , 1987, Cell.

[32]  H. Riezman,et al.  Determinants for glycophospholipid anchoring of the Saccharomyces cerevisiae GAS1 protein to the plasma membrane , 1991, Molecular and cellular biology.

[33]  R. Lefkowitz,et al.  GTPase Activating Specificity of RGS12 and Binding Specificity of an Alternatively Spliced PDZ (PSD-95/Dlg/ZO-1) Domain* , 1998, The Journal of Biological Chemistry.

[34]  I. Herskowitz,et al.  Functional analysis of the interaction between the small GTP binding protein Cdc42 and the Ste20 protein kinase in yeast. , 1996, The EMBO journal.

[35]  Necmettin Yildirim,et al.  Regulators of G Protein Signaling and Transient Activation of Signaling , 2003, Journal of Biological Chemistry.

[36]  M. Seedorf,et al.  Asc1p, a WD40-domain containing adaptor protein, is required for the interaction of the RNA-binding protein Scp160p with polysomes. , 2004, The Biochemical journal.

[37]  J. Hepler,et al.  Cellular Regulation of RGS Proteins: Modulators and Integrators of G Protein Signaling , 2002, Pharmacological Reviews.

[38]  M. Whiteway,et al.  The protein kinase homologue Ste20p is required to link the yeast pheromone response G‐protein beta gamma subunits to downstream signalling components. , 1992, The EMBO journal.

[39]  H. Kitano,et al.  A quantitative characterization of the yeast heterotrimeric G protein cycle , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[40]  J. Thevelein,et al.  Glucose and sucrose act as agonist and mannose as antagonist ligands of the G protein-coupled receptor Gpr1 in the yeast Saccharomyces cerevisiae. , 2004, Molecular cell.

[41]  J. Hirsch,et al.  Krh1p and Krh2p act downstream of the Gpa2p Gα subunit to negatively regulate haploid invasive growth , 2003, Journal of Cell Science.

[42]  J. D. de Winde,et al.  A Saccharomyces cerevisiae G‐protein coupled receptor, Gpr1, is specifically required for glucose activation of the cAMP pathway during the transition to growth on glucose , 1999, Molecular microbiology.

[43]  J. Thorner,et al.  Pheromone action regulates G-protein alpha-subunit myristoylation in the yeast Saccharomyces cerevisiae. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[44]  H. Horvitz,et al.  EGL-10 Regulates G Protein Signaling in the C. elegans Nervous System and Shares a Conserved Domain with Many Mammalian Proteins , 1996, Cell.

[45]  S. Reed,et al.  Role for the Rho-family GTPase Cdc42 in yeast mating-pheromone signal pathway , 1995, Nature.

[46]  G K Lewis,et al.  Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product , 1985, Molecular and cellular biology.

[47]  M. Lemmon,et al.  All Phox Homology (PX) Domains from Saccharomyces cerevisiae Specifically Recognize Phosphatidylinositol 3-Phosphate* , 2001, The Journal of Biological Chemistry.

[48]  K. Arai,et al.  Regulation of the yeast pheromone response pathway by G protein subunits. , 1990, The EMBO journal.

[49]  A. Gilman,et al.  RGS4 and GAIP are GTPase-activating proteins for Gq alpha and block activation of phospholipase C beta by gamma-thio-GTP-Gq alpha. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[50]  M. Dong,et al.  Multiple RGS proteins alter neural G protein signaling to allow C. elegans to rapidly change behavior when fed. , 2000, Genes & development.

[51]  M. Snyder,et al.  A genomic study of the bipolar bud site selection pattern in Saccharomyces cerevisiae. , 2001, Molecular biology of the cell.

[52]  M. Peter,et al.  Actin cytoskeleton organization regulated by the PAK family of protein kinases , 1998, Current Biology.

[53]  A. Hopkins,et al.  The druggable genome , 2002, Nature Reviews Drug Discovery.

[54]  J. François,et al.  Involvement of GFA1, which encodes glutamine-fructose-6-phosphate amidotransferase, in the activation of the chitin synthesis pathway in response to cell-wall defects in Saccharomyces cerevisiae. , 2002, European journal of biochemistry.

[55]  R. K. Chan,et al.  Isolation and genetic analysis of Saccharomyces cerevisiae mutants supersensitive to G1 arrest by a factor and alpha factor pheromones , 1982, Molecular and cellular biology.

[56]  J. D. de Winde,et al.  A novel regulator of G protein signalling in yeast, Rgs2, downregulates glucose‐activation of the cAMP pathway through direct inhibition of Gpa2 , 1999, The EMBO journal.

[57]  H. Dohlman,et al.  Endoproteolytic Processing of Sst2, a Multidomain Regulator of G Protein Signaling in Yeast* , 2000, The Journal of Biological Chemistry.

[58]  E. Ross,et al.  Phospholipase C-β1 is a GTPase-activating protein for Gq/11, its physiologic regulator , 1992, Cell.

[59]  R. K. Chan,et al.  Physiological characterization of Saccharomyces cerevisiae mutants supersensitive to G1 arrest by a factor and alpha factor pheromones , 1982, Molecular and cellular biology.

[60]  C. Carman,et al.  Selective Regulation of Gαq/11 by an RGS Domain in the G Protein-coupled Receptor Kinase, GRK2* , 1999, The Journal of Biological Chemistry.

[61]  Richard R. Neubig,et al.  Regulators of G-Protein signalling as new central nervous system drug targets , 2002, Nature Reviews Drug Discovery.

[62]  E. Winter,et al.  An osmosensing signal transduction pathway in yeast. , 1993, Science.

[63]  H. Dohlman,et al.  Regulation of Membrane and Subunit Interactions by N-Myristoylation of a G Protein α Subunit in Yeast* , 1996, The Journal of Biological Chemistry.

[64]  Liam J. McGuffin,et al.  The PSIPRED protein structure prediction server , 2000, Bioinform..

[65]  Daria Mochly-Rosen,et al.  Adaptor proteins in protein kinase C-mediated signal transduction , 2001, Oncogene.

[66]  A. Wittinghofer.  Signaling mechanistics: Aluminum fluoride for molecule of the year , 1997, Current Biology.

[67]  Mariusz Olczak,et al.  Chitin Synthesis in Saccharomyces cerevisiae in Response to Supplementation of Growth Medium with Glucosamine and Cell Wall Stress , 2003, Eukaryotic Cell.

[68]  J. Thorner,et al.  Regulation of G protein-initiated signal transduction in yeast: paradigms and principles. , 2001, Annual review of biochemistry.

[69]  P B Sigler,et al.  Sst2 is a GTPase-activating protein for Gpa1: purification and characterization of a cognate RGS-Galpha protein pair in yeast. , 1998, Biochemistry.

[70]  K. Tang,et al.  Spatial and Temporal Regulation of RACK1 Function and N-methyl-D-aspartate Receptor Activity through WD40 Motif-mediated Dimerization* , 2004, Journal of Biological Chemistry.

[71]  H. Hamm,et al.  Interaction of Gβγ with RACK1 and other WD40 repeat proteins , 2004 .

[72]  D. Drubin,et al.  Spontaneous receptor-independent heterotrimeric G-protein signalling in an RGS mutant , 2003, Nature Cell Biology.

[73]  K Nasmyth,et al.  growth and for cytokinesis in budding yeast. Ste20-like protein kinases are required for normal localization of cell , 2007 .

[74]  J. Armstrong,et al.  Characterization of the Bar proteinase, an extracellular enzyme from the yeast Saccharomyces cerevisiae. , 1991, Advances in experimental medicine and biology.

[75]  M. Whiteway,et al.  Overexpression of the STE4 gene leads to mating response in haploid Saccharomyces cerevisiae. , 1990, Molecular and cellular biology.

[76]  I. Herskowitz,et al.  The Hog1 MAPK prevents cross talk between the HOG and pheromone response MAPK pathways in Saccharomyces cerevisiae. , 1998, Genes & development.

[77]  Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product , 1985 .

[78]  J. Fridovich-Keil,et al.  Scp160p, a multiple KH-domain protein, is a component of mRNP complexes in yeast. , 2000, Nucleic acids research.

[79]  J. Jordan,et al.  Regulators of G Protein Signaling Proteins as Determinants of the Rate of Desensitization of Presynaptic Calcium Channels* , 1999, The Journal of Biological Chemistry.

[80]  J. Thorner,et al.  A presumptive helicase (MOT1 gene product) affects gene expression and is required for viability in the yeast Saccharomyces cerevisiae , 1992, Molecular and cellular biology.

[81]  J. Thompson,et al.  The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. , 1997, Nucleic acids research.

[82]  J. Heitman,et al.  The Gα Protein Gpa2 Controls Yeast Differentiation by Interacting with Kelch Repeat Proteins that Mimic Gβ Subunits , 2002 .

[83]  S. Emr,et al.  Phox domain interaction with PtdIns(3)P targets the Vam7 t-SNARE to vacuole membranes , 2001, Nature Cell Biology.

[84]  W. Weis,et al.  Structural basis of the Axin–adenomatous polyposis coli interaction , 2000, The EMBO journal.

[85]  M. Sallese,et al.  Selective regulation of Gq signaling by G protein-coupled receptor kinase 2: direct interaction of kinase N terminus with activated galphaq. , 2000, Molecular pharmacology.

[86]  M. Nakafuku,et al.  Occurrence in Saccharomyces cerevisiae of a gene homologous to the cDNA coding for the alpha subunit of mammalian G proteins. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[87]  L. Lim,et al.  Pheromone signalling in Saccharomyces cerevisiae requires the small GTP-binding protein Cdc42p and its activator CDC24 , 1995, Molecular and cellular biology.

[88]  A. Gilman,et al.  The GTPase-activating Protein RGS4 Stabilizes the Transition State for Nucleotide Hydrolysis* , 1996, The Journal of Biological Chemistry.

[89]  H. Dohlman,et al.  Identification of yeast pheromone pathway modulators by high-throughput agonist response profiling of a yeast gene knockout strain collection. , 2004, Methods in enzymology.

[90]  H. Dohlman,et al.  G proteins and pheromone signaling. , 2002, Annual review of physiology.

[91]  M. Tyers,et al.  A new family of regulators of G-protein-coupled receptors? , 1996, Current Biology.

[92]  Howard Riezman,et al.  Ubiquitination of a Yeast Plasma Membrane Receptor Signals Its Ligand-Stimulated Endocytosis , 1996, Cell.

[93]  S. Reed,et al.  Stoichiometry of G protein subunits affects the Saccharomyces cerevisiae mating pheromone signal transduction pathway , 1990, Molecular and cellular biology.

[94]  J. Benovic,et al.  G protein-coupled receptor Kinase 2/G alpha q/11 interaction. A novel surface on a regulator of G protein signaling homology domain for binding G alpha subunits. , 2003, The Journal of biological chemistry.

[95]  S. Popov,et al.  Determination of the Contact Energies between a Regulator of G Protein Signaling and G Protein Subunits and Phospholipase Cβ1 , 2001 .

[96]  G F Sprague,et al.  Mutation of RGA1, which encodes a putative GTPase-activating protein for the polarity-establishment protein Cdc42p, activates the pheromone-response pathway in the yeast Saccharomyces cerevisiae. , 1995, Genes & development.

[97]  M. Snyder,et al.  Molecular basis of cell integrity and morphogenesis in Saccharomyces cerevisiae. , 1995, Microbiological reviews.

[98]  J. Thevelein,et al.  Sex and sugar in yeast: two distinct GPCR systems , 2001, EMBO reports.

[99]  H. Hamm,et al.  RACK1 Regulates Specific Functions of Gβγ* , 2004, Journal of Biological Chemistry.

[100]  N. Grishin,et al.  Evolution of the regulators of G-protein signaling multigene family in mouse and human. , 2002, Genomics.

[101]  C. Dessauer,et al.  Regulator of G-protein Signaling 3 (RGS3) Inhibits Gβ1γ2-induced Inositol Phosphate Production, Mitogen-activated Protein Kinase Activation, and Akt Activation* , 2001, The Journal of Biological Chemistry.

[102]  M. Yaffe,et al.  Temperature-sensitive yeast mutants defective in mitochondrial inheritance , 1990, The Journal of cell biology.

[103]  I. Herskowitz,et al.  From membrane to nucleus: the pathway of signal transduction in yeast and its genetic control. , 1988, Cold Spring Harbor symposia on quantitative biology.

[104]  M. Valdivieso,et al.  Chitin Synthesis in a gas1 Mutant ofSaccharomyces cerevisiae , 2000, Journal of bacteriology.

[105]  C. Lions,et al.  The transcriptional regulator Hap1p (Cyp1p) is essential for anaerobic or heme-deficient growth of Saccharomyces cerevisiae: Genetic and molecular characterization of an extragenic suppressor that encodes a WD repeat protein. , 1998, Genetics.

[106]  P. Sternberg,et al.  Antagonism between Goα and Gqα in Caenorhabditis elegans: the RGS protein EAT-16 is necessary for Goα signaling and regulates Gqα activity , 1999 .

[107]  J. Chant,et al.  Subcellular Localization of Axl1, the Cell Type-Specific Regulator of Polarity , 2002, Current Biology.

[108]  K. Blumer,et al.  RGS family members: GTPase-activating proteins for heterotrimeric G-protein α-subunits , 1996, Nature.

[109]  G. Müller,et al.  A cAMP-binding ectoprotein in the yeast Saccharomyces cerevisiae. , 1991, Biochemistry.

[110]  H. Hamm,et al.  Crystal structure of a G-protein beta gamma dimer at 2.1A resolution. , 1996, Nature.

[111]  J. Thorner,et al.  Sst2, a negative regulator of pheromone signaling in the yeast Saccharomyces cerevisiae: expression, localization, and genetic interaction and physical association with Gpa1 (the G-protein alpha subunit) , 1996, Molecular and cellular biology.

[112]  Kevan M. Shokat,et al.  Chemical genetic analysis of the budding-yeast p21-activated kinase Cla4p , 2000, Nature Cell Biology.

[113]  H. Dohlman,et al.  Selective Uncoupling of RGS Action by a Single Point Mutation in the G Protein α-Subunit* , 1998, The Journal of Biological Chemistry.

[114]  P. Brown,et al.  Mediator protein mutations that selectively abolish activated transcription. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[115]  H. Dohlman,et al.  Inhibition of G-protein signaling by dominant gain-of-function mutations in Sst2p, a pheromone desensitization factor in Saccharomyces cerevisiae , 1995, Molecular and cellular biology.

[116]  J. D. de Winde,et al.  Glucose‐induced cAMP signalling in yeast requires both a G‐protein coupled receptor system for extracellular glucose detection and a separable hexose kinase‐dependent sensing process , 2000, Molecular microbiology.

[117]  Thierry Fontaine,et al.  Glycosylphosphatidylinositol-anchored Glucanosyltransferases Play an Active Role in the Biosynthesis of the Fungal Cell Wall* , 2000, The Journal of Biological Chemistry.

[118]  P. Bonfante,et al.  Increase in chitin as an essential response to defects in assembly of cell wall polymers in the ggp1delta mutant of Saccharomyces cerevisiae , 1997, Journal of bacteriology.

[119]  Joseph D. Schrag,et al.  Interaction of a G-protein β-subunit with a conserved sequence in Ste20/PAK family protein kinases , 1998, Nature.

[120]  K. Arai,et al.  GPA1, a haploid-specific essential gene, encodes a yeast homolog of mammalian G protein which may be involved in mating factor signal transduction , 1987, Cell.

[121]  G. Sprague,,et al.  The Phosphotyrosyl Phosphatase Activator, Ncs1p (Rrd1p), Functions with Cla4p To Regulate the G2/M Transition inSaccharomyces cerevisiae , 2001, Molecular and Cellular Biology.

[122]  Ronald W. Davis,et al.  A genome-wide screen in Saccharomyces cerevisiae for genes affecting UV radiation sensitivity , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[123]  Hidekazu Hiroaki,et al.  Solution structure of the PX domain, a target of the SH3 domain , 2001, Nature Structural Biology.

[124]  Lawrence Lum,et al.  Identification of Hedgehog Pathway Components by RNAi in Drosophila Cultured Cells , 2003, Science.

[125]  M. Yaffe,et al.  Mutational Analysis of Mdm1p Function in Nuclear and Mitochondrial Inheritance , 1997, The Journal of cell biology.

[126]  Prahlad T. Ram,et al.  G Protein Pathways , 2002, Science.

[127]  P. H. Anborgh,et al.  G Protein-coupled Receptor Kinase 2 Regulator of G Protein Signaling Homology Domain Binds to Both Metabotropic Glutamate Receptor 1a and Gαq to Attenuate Signaling* , 2004, Journal of Biological Chemistry.

[128]  J. Chant,et al.  Rax1, a protein required for the establishment of the bipolar budding pattern in yeast. , 2004, Gene.

[129]  David Y. Thomas,et al.  The STE4 and STE18 genes of yeast encode potential β and γ subunits of the mating factor receptor-coupled G protein , 1989, Cell.

[130]  J. Yates,et al.  Direct analysis of protein complexes using mass spectrometry , 1999, Nature Biotechnology.

[131]  F. Klis,et al.  Cell wall perturbation in yeast results in dual phosphorylation of the Slt2/Mpk1 MAP kinase and in an Slt2-mediated increase in FKS2-lacZ expression, glucanase resistance and thermotolerance. , 2000, Microbiology.

[132]  J. Hoheisel,et al.  Genome-wide Analysis of the Response to Cell Wall Mutations in the Yeast Saccharomyces cerevisiae* , 2003, Journal of Biological Chemistry.

[133]  G. Sprague,,et al.  Assay of yeast mating reaction. , 1991, Methods in enzymology.

[134]  A. Gilman,et al.  GAIP and RGS4 Are GTPase-Activating Proteins for the Gi Subfamily of G Protein α Subunits , 1996, Cell.

[135]  C. Dessauer,et al.  RGS2 regulates signal transduction in olfactory neurons by attenuating activation of adenylyl cyclase III , 2001, Nature.

[136]  M. Valdivieso,et al.  Maintenance of cell integrity in the gas1 mutant of Saccharomyces cerevisiae requires the Chs3p‐targeting and activation pathway and involves an unusual Chs3p localization , 2002, Yeast.

[137]  N. Tjandra,et al.  Solution structure of human GAIP (Galpha interacting protein): a regulator of G protein signaling. , 1999, Journal of molecular biology.

[138]  E. Elion,et al.  The osmoregulatory pathway represses mating pathway activity in Saccharomyces cerevisiae: isolation of a FUS3 mutant that is insensitive to the repression mechanism , 1996, Molecular and cellular biology.

[139]  Nancy Kleckner,et al.  A Method for Gene Disruption That Allows Repeated Use of URA3 Selection in the Construction of Multiply Disrupted Yeast Strains , 1987, Genetics.

[140]  S. Grill,et al.  RGS-7 Completes a Receptor-Independent Heterotrimeric G Protein Cycle to Asymmetrically Regulate Mitotic Spindle Positioning in C. elegans , 2004, Cell.