14-3-3 Proteins Are Essential for RAS/MAPK Cascade Signaling during Pseudohyphal Development in S. cerevisiae

14-3-3 proteins are highly conserved ubiquitous proteins whose explicit functions have remained elusive. Here, we show that the S. cerevisiae 14-3-3 homologs BMH1 and BMH2 are not essential for viability or mating MAPK cascade signaling, but they are essential for pseudohyphal-development MAPK cascade signaling and other processes. Activated alleles of RAS2 and CDC42 induce pseudohyphal development and FG(TyA)-lacZ signaling in Bmh+ strains but not in ste20 (p65PAK) or bmh1 bmh2 mutant strains. Moreover, Bmh1p and Bmh2p associate with Ste20p in vivo. Three alleles of BMH1 encode proteins defective for FG(TyA)-lacZ signaling and association with Ste20p, yet these alleles complement other 14-3-3 functions. Therefore, the 14-3-3 proteins are specifically required for RAS/MAPK cascade signaling during pseudohyphal development in S. cerevisiae.

[1]  P. Silver,et al.  The GTP-bound form of the yeast Ran/TC4 homologue blocks nuclear protein import and appearance of poly(A)+ RNA in the cytoplasm. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[2]  G. Fink,et al.  Elements of the yeast pheromone response pathway required for filamentous growth of diploids. , 1993, Science.

[3]  T. Isobe,et al.  Identification of the Site of Interaction of the 14-3-3 Protein with Phosphorylated Tryptophan Hydroxylase (*) , 1995, The Journal of Biological Chemistry.

[4]  D. Morrison,et al.  14-3-3 is not essential for Raf-1 function: identification of Raf-1 proteins that are biologically activated in a 14-3-3- and Ras-independent manner , 1995, Molecular and cellular biology.

[5]  C. J. Gimeno,et al.  Saccharomyces cerevisiae TEC1 is required for pseudohyphal growth , 1996, Molecular microbiology.

[6]  P. Allen,et al.  Interaction of 14-3-3 with Signaling Proteins Is Mediated by the Recognition of Phosphoserine , 1996, Cell.

[7]  R. Ferl,et al.  Four Arabidopsis thaliana 14‐3‐3 protein isoforms can complement the lethal yeast bmh1 bmh2 double disruption , 1996, FEBS letters.

[8]  R. Burgoyne,et al.  Distinct effects of alpha-SNAP, 14-3-3 proteins, and calmodulin on priming and triggering of regulated exocytosis , 1995, The Journal of cell biology.

[9]  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.

[10]  G. Fink,et al.  Elements of a single MAP kinase cascade in Saccharomyces cerevisiae mediate two developmental programs in the same cell type: mating and invasive growth. , 1994, Genes & development.

[11]  Gerald R. Fink,et al.  Unipolar cell divisions in the yeast S. cerevisiae lead to filamentous growth: Regulation by starvation and RAS , 1992, Cell.

[12]  C. Kaiser,et al.  Yeast SEC16 gene encodes a multidomain vesicle coat protein that interacts with Sec23p , 1995, The Journal of cell biology.

[13]  A. Aitken,et al.  14-3-3 proteins : a highly conserved widespread family of eukaryotic proteins , 2003 .

[14]  Michael Wigler,et al.  Three different genes in S. cerevisiae encode the catalytic subunits of the cAMP-dependent protein kinase , 1987, Cell.

[15]  J. Chant,et al.  Rac and Cdc42 Induce Actin Polymerization and G1 Cell Cycle Progression Independently of p65PAK and the JNK/SAPK MAP Kinase Cascade , 1996, Cell.

[16]  J. Segall,et al.  Functional characterization of the Cdc42p binding domain of yeast Ste20p protein kinase , 1997, The EMBO journal.

[17]  S. Smerdon,et al.  Structure of a 14-3-3 protein and implications for coordination of multiple signalling pathways , 1995, Nature.

[18]  K. Irie,et al.  Stimulatory effects of yeast and mammalian 14-3-3 proteins on the Raf protein kinase. , 1994, Science.

[19]  G. Fink,et al.  Dissection of filamentous growth by transposon mutagenesis in Saccharomyces cerevisiae. , 1997, Genetics.

[20]  B. Cairns,et al.  Order of action of components in the yeast pheromone response pathway revealed with a dominant allele of the STE11 kinase and the multiple phosphorylation of the STE7 kinase. , 1992, Genes & development.

[21]  K. Irie,et al.  14-3-3 proteins: potential roles in vesicular transport and Ras signaling in Saccharomyces cerevisiae. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[22]  A Aitken,et al.  Isoforms of 14‐3‐3 protein can form homo‐ and heterodimers in vivo and in vitro: implications for function as adapter proteins , 1995, FEBS letters.

[23]  G. Fink,et al.  Combinatorial Control Required for the Specificity of Yeast MAPK Signaling , 1997, Science.

[24]  I. Herskowitz.  MAP kinase pathways in yeast: For mating and more , 1995, Cell.

[25]  B. Errede,et al.  Pheromone-induced signal transduction in Saccharomyces cerevisiae requires the sequential function of three protein kinases , 1993, Molecular and cellular biology.

[26]  R. Liddington,et al.  Crystal structure of the zeta isoform of the 14-3-3 protein , 1995, Nature.

[27]  J. Thorner,et al.  12 Pheromone Response and Signal Transduction during the Mating Process of Saccharomyces cerevisiae , 1992 .

[28]  S. Fields,et al.  Overproduction of the yeast STE12 protein leads to constitutive transcriptional induction. , 1990, Genes & development.

[29]  E. Dubois,et al.  Involvement of SRE element of Ty1 transposon in TEC1-dependent transcriptional activation. , 1994, Nucleic acids research.

[30]  M. Barbacid,et al.  Lack of evidence for the activation of the Ras/Raf mitogenic pathway by 14-3-3 proteins in mammalian cells. , 1995, Oncogene.

[31]  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.

[32]  A. Carr,et al.  14-3-3 protein homologs required for the DNA damage checkpoint in fission yeast. , 1994, Science.

[33]  W. Kolch,et al.  Regulation of Raf‐1 kinase activity by the 14‐3‐3 family of proteins. , 1995, The EMBO journal.

[34]  F. McCormick,et al.  Binding of 14-3-3 proteins to the protein kinase Raf and effects on its activation. , 1994, Science.

[35]  G. Fink,et al.  Methods in yeast genetics , 1979 .

[36]  A. Toker,et al.  Multiple isoforms of a protein kinase C inhibitor (KCIP-1/14-3-3) from sheep brain. Amino acid sequence of phosphorylated forms. , 1992, European journal of biochemistry.

[37]  G. Fink,et al.  Ras2 signals via the Cdc42/Ste20/mitogen-activated protein kinase module to induce filamentous growth in Saccharomyces cerevisiae. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[38]  W. Fantl,et al.  Activation of Raf-1 by 14-3-3 proteins , 1994, Nature.

[39]  L. Giot,et al.  Functional analysis of the interaction between Afr1p and the Cdc12p septin, two proteins involved in pheromone-induced morphogenesis. , 1997, Molecular biology of the cell.

[40]  K. Xia,et al.  Interaction of the protein kinase Raf-1 with 14-3-3 proteins. , 1994, Science.

[41]  M. Wigler,et al.  cAMP-independent control of sporulation, glycogen metabolism, and heat shock resistance in S. cerevisiae , 1988, Cell.