The TEA Transcription Factor Tec1 Links TOR and MAPK Pathways to Coordinate Yeast Development

In Saccharomyces cerevisiae, the TEA transcription factor Tec1 controls several developmental programs in response to nutrients and pheromones. Tec1 is targeted by the pheromone-responsive Fus3/Kss1 mitogen-activated protein kinase (MAPK) cascade, which destabilizes the transcription factor to ensure efficient mating of sexual partner cells. The regulation of Tec1 by signaling pathways that control cell division and development in response to nutrients, however, is not known. Here, we show that Tec1 protein stability is under control of the nutrient-sensitive target of rapamycin complex 1 (TORC1) signaling pathway via the Tip41-Tap42-Sit4 branch. We further show that degradation of Tec1 upon inhibition of TORC1 by rapamycin does not involve polyubiquitylation and appears to be proteasome independent. However, rapamycin-induced Tec1 degradation depends on the HECT ubiquitin ligase Rsp5, which physically interacts with Tec1 via conserved PxY motives. We further demonstrate that rapamycin and mating pheromone control Tec1 protein stability through distinct mechanisms by targeting different domains of the transcription factor. Finally, we show that Tec1 is a positive regulator of yeast chronological lifespan (CLS), a known TORC1-regulated process. Our findings indicate that in yeast, Tec1 links TORC1 and MAPK signaling pathways to coordinate control of cellular development in response to different stimuli.

[1]  W. Timberlake,et al.  ATTS, a new and conserved DNA binding domain. , 1991, The Plant cell.

[2]  Lan Huang,et al.  Fus3-Regulated Tec1 Degradation through SCFCdc4 Determines MAPK Signaling Specificity during Mating in Yeast , 2004, Cell.

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

[4]  D. Wolf,et al.  Yeast cycloheximide-resistant crl mutants are proteasome mutants defective in protein degradation. , 1997, Molecular biology of the cell.

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

[6]  G. Braus,et al.  Asymmetrically localized Bud8p and Bud9p proteins control yeast cell polarity and development , 2000, The EMBO journal.

[7]  D. Lane,et al.  Updates on p53: modulation of p53 degradation as a therapeutic approach , 2008, British Journal of Cancer.

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

[9]  Curt Wittenberg,et al.  G1-specific cyclins of S. cerevisiae: Cell cycle periodicity, regulation by mating pheromone, and association with the p34 CDC28 protein kinase , 1990, Cell.

[10]  P. Kogut,et al.  Protein kinase-A dependent phosphorylation of transcription enhancer factor-1 represses its DNA-binding activity but enhances its gene activation ability. , 2000, Nucleic acids research.

[11]  R. D. Gietz,et al.  New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. , 1988, Gene.

[12]  Saeed Tavazoie,et al.  Ras and Gpa2 Mediate One Branch of a Redundant Glucose Signaling Pathway in Yeast , 2004, PLoS biology.

[13]  B. André,et al.  The Npr1 Kinase Controls Biosynthetic and Endocytic Sorting of the Yeast Gap1 Permease* , 2001, The Journal of Biological Chemistry.

[14]  J. Broach,et al.  The role of phosphatases in TOR signaling in yeast. , 2004, Current topics in microbiology and immunology.

[15]  K. Arndt,et al.  TIP41 interacts with TAP42 and negatively regulates the TOR signaling pathway. , 2001, Molecular cell.

[16]  Lee Bardwell,et al.  A walk-through of the yeast mating pheromone response pathway , 2004, Peptides.

[17]  R. Hay.  Role of ubiquitin-like proteins in transcriptional regulation. , 2006, Ernst Schering Research Foundation workshop.

[18]  N. Ogawa,et al.  A series of double disruptants for protein phosphatase genes in Saccharomyces cerevisiae and their phenotypic analysis , 2002, Yeast.

[19]  R. Sikorski,et al.  A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. , 1989, Genetics.

[20]  A. Toh-E,et al.  The PY-motif of Bul1 protein is essential for growth of Saccharomyces cerevisiae under various stress conditions. , 1998, Gene.

[21]  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.

[22]  R. Hay,et al.  Regulation of transcription factors by protein degradation , 2000, Cellular and Molecular Life Sciences CMLS.

[23]  P. Polak,et al.  mTOR and the control of whole body metabolism. , 2009, Current opinion in cell biology.

[24]  R. Loewith,et al.  Cell growth control: little eukaryotes make big contributions , 2006, Oncogene.

[25]  Stefan Brückner,et al.  Differential regulation of Tec1 by Fus3 and Kss1 confers signaling specificity in yeast development , 2004, Current Genetics.

[26]  James R Broach,et al.  How Saccharomyces responds to nutrients. , 2008, Annual review of genetics.

[27]  H. Mösch,et al.  The TEA Transcription Factor Tec1 Confers Promoter-Specific Gene Regulation by Ste12-Dependent and -Independent Mechanisms , 2010, Eukaryotic Cell.

[28]  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.

[29]  E. Golemis,et al.  LexA-based two-hybrid systems. , 2000, Methods in enzymology.

[30]  Shelley Lane,et al.  Regulation of Mating and Filamentation Genes by Two Distinct Ste12 Complexes in Saccharomyces cerevisiae , 2006, Molecular and Cellular Biology.

[31]  J. Heitman,et al.  Protein Kinase A Operates a Molecular Switch That Governs Yeast Pseudohyphal Differentiation , 2002, Molecular and Cellular Biology.

[32]  V. Longo,et al.  Regulation of Longevity and Stress Resistance by Sch9 in Yeast , 2001, Science.

[33]  Chao Cheng,et al.  Life Span Extension by Calorie Restriction Depends on Rim15 and Transcription Factors Downstream of Ras/PKA, Tor, and Sch9 , 2007, PLoS genetics.

[34]  S. Jentsch,et al.  Activation of a Membrane-Bound Transcription Factor by Regulated Ubiquitin/Proteasome-Dependent Processing , 2000, Cell.

[35]  J. Heitman,et al.  The TOR signal transduction cascade controls cellular differentiation in response to nutrients. , 2001, Molecular biology of the cell.

[36]  A. Gruhler,et al.  PRE2, highly homologous to the human major histocompatibility complex-linked RING10 gene, codes for a yeast proteasome subunit necessary for chrymotryptic activity and degradation of ubiquitinated proteins. , 1993, The Journal of biological chemistry.

[37]  Matt Kaeberlein,et al.  Extension of chronological life span in yeast by decreased TOR pathway signaling. , 2006, Genes & development.

[38]  A. Kikuchi,et al.  PY motifs of Rod1 are required for binding to Rsp5 and for drug resistance , 2002, FEBS letters.

[39]  V. Longo.  The Ras and Sch9 pathways regulate stress resistance and longevity , 2003, Experimental Gerontology.

[40]  J. Huibregtse,et al.  The Rsp5 ubiquitin ligase is coupled to and antagonized by the Ubp2 deubiquitinating enzyme , 2005, The EMBO journal.

[41]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[42]  P. Nurse,et al.  TOR signalling regulates mitotic commitment through the stress MAP kinase pathway and the Polo and Cdc2 kinases , 2007, Nature Cell Biology.

[43]  N. Eberhardt,et al.  DNA Binding of TEA/ATTS Domain Factors Is Regulated by Protein Kinase C Phosphorylation in Human Choriocarcinoma Cells* , 2001, The Journal of Biological Chemistry.

[44]  Robbie Loewith,et al.  Caffeine extends yeast lifespan by targeting TORC1 , 2008, Molecular microbiology.

[45]  J. Crespo,et al.  NPR1 Kinase and RSP5-BUL1/2 Ubiquitin Ligase Control GLN3-dependent Transcription in Saccharomyces cerevisiae* , 2004, Journal of Biological Chemistry.

[46]  I. Simon,et al.  Program-Specific Distribution of a Transcription Factor Dependent on Partner Transcription Factor and MAPK Signaling , 2003, Cell.

[47]  A. Schmidt,et al.  The TOR nutrient signalling pathway phosphorylates NPR1 and inhibits turnover of the tryptophan permease , 1998, The EMBO journal.

[48]  G. Fink,et al.  Crosstalk between the Ras2p-controlled mitogen-activated protein kinase and cAMP pathways during invasive growth of Saccharomyces cerevisiae. , 1999, Molecular biology of the cell.

[49]  R. Klein,et al.  TEF-1 and C/EBPbeta are major p38alpha MAPK-regulated transcription factors in proliferating cardiomyocytes. , 2006, The Biochemical journal.

[50]  K. Arndt,et al.  The SAP, a new family of proteins, associate and function positively with the SIT4 phosphatase , 1996, Molecular and cellular biology.

[51]  J. Huibregtse,et al.  A Single PXY Motif Located within the Carboxyl Terminus of Spt23p and Mga2p Mediates a Physical and Functional Interaction with Ubiquitin Ligase Rsp5p* , 2004, Journal of Biological Chemistry.

[52]  A. Davies,et al.  Activation of Ubiquitin-Dependent DNA Damage Bypass Is Mediated by Replication Protein A , 2008, Molecular cell.

[53]  V. Kushnirov.  Rapid and reliable protein extraction from yeast , 2000, Yeast.

[54]  Robbie Loewith,et al.  Sch9 is a major target of TORC1 in Saccharomyces cerevisiae. , 2007, Molecular cell.

[55]  Serguei Sokol,et al.  Investigating the caffeine effects in the yeast Saccharomyces cerevisiae brings new insights into the connection between TOR, PKC and Ras/cAMP signalling pathways , 2006, Molecular microbiology.

[56]  John R. Yates,et al.  Pheromone-Dependent Destruction of the Tec1 Transcription Factor Is Required for MAP Kinase Signaling Specificity in Yeast , 2004, Cell.

[57]  Henrik G. Dohlman,et al.  Pheromone-regulated Sumoylation of Transcription Factors That Mediate the Invasive to Mating Developmental Switch in Yeast* , 2006, Journal of Biological Chemistry.

[58]  T. Boller,et al.  Saccharomyces cerevisiae cAMP-dependent protein kinase controls entry into stationary phase through the Rim15p protein kinase. , 1998, Genes & development.

[59]  M. Cárdenas,et al.  Nutritional control via Tor signaling in Saccharomyces cerevisiae. , 2008, Current opinion in microbiology.

[60]  W. Lo,et al.  The cell surface flocculin Flo11 is required for pseudohyphae formation and invasion by Saccharomyces cerevisiae. , 1998, Molecular biology of the cell.

[61]  J. Heitman,et al.  Targets for cell cycle arrest by the immunosuppressant rapamycin in yeast , 1991, Science.

[62]  G. Fink,et al.  The three yeast A kinases have specific signaling functions in pseudohyphal growth. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[63]  E. Herrero,et al.  Sit4 Is Required for Proper Modulation of the Biological Functions Mediated by Pkc1 and the Cell Integrity Pathway inSaccharomyces cerevisiae * , 2002, The Journal of Biological Chemistry.

[64]  A. Davies,et al.  Ubiquitylation of the 9-1-1 Checkpoint Clamp Is Independent of Rad6-Rad18 and DNA Damage , 2010, Cell.

[65]  I. Stansfield,et al.  An MBoC Favorite: TOR controls translation initiation and early G1 progression in yeast , 2012, Molecular biology of the cell.

[66]  Lilia Alberghina,et al.  Rapamycin‐mediated G1 arrest involves regulation of the Cdk inhibitor Sic1 in Saccharomyces cerevisiae , 2007, Molecular microbiology.

[67]  Matt Kaeberlein,et al.  Regulation of Yeast Replicative Life Span by TOR and Sch9 in Response to Nutrients , 2005, Science.

[68]  Ivo Pedruzzi,et al.  TOR and PKA signaling pathways converge on the protein kinase Rim15 to control entry into G0. , 2003, Molecular cell.

[69]  M. Ward,et al.  Yeast PKA represses Msn2p/Msn4p‐dependent gene expression to regulate growth, stress response and glycogen accumulation , 1998, The EMBO journal.

[70]  N. Gueven,et al.  The complexity of p53 stabilization and activation , 2006, Cell Death and Differentiation.

[71]  R. Klein,et al.  TEF-1 and C/EBPβ are major p38α MAPK-regulated transcription factors in proliferating cardiomyocytes , 2006 .

[72]  E. Lander,et al.  Effectors of a developmental mitogen-activated protein kinase cascade revealed by expression signatures of signaling mutants. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[73]  F. Winston,et al.  A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. , 1987, Gene.

[74]  Roger Brent,et al.  C dil, a Human Gl and S Phase Protein Phosphatase That Associates with Cdk2 , 2003 .

[75]  D. Rotin,et al.  Physiological functions of the HECT family of ubiquitin ligases , 2009, Nature Reviews Molecular Cell Biology.

[76]  Xiaolian Gao,et al.  Insights into transcription enhancer factor 1 (TEF-1) activity from the solution structure of the TEA domain , 2006, Proceedings of the National Academy of Sciences.

[77]  Gerald R. Fink,et al.  MAP Kinases with Distinct Inhibitory Functions Impart Signaling Specificity during Yeast Differentiation , 1997, Cell.

[78]  Gerald R. Fink,et al.  Guide to yeast genetics and molecular biology , 1993 .

[79]  Michael N. Hall,et al.  Elucidating TOR Signaling and Rapamycin Action: Lessons from Saccharomyces cerevisiae , 2002, Microbiology and Molecular Biology Reviews.

[80]  J. Thorner,et al.  Function and regulation in MAPK signaling pathways: lessons learned from the yeast Saccharomyces cerevisiae. , 2007, Biochimica et biophysica acta.

[81]  H. Tamaki.  Glucose-stimulated cAMP-protein kinase A pathway in yeast Saccharomyces cerevisiae. , 2007, Journal of bioscience and bioengineering.

[82]  G. Braus,et al.  Dual Role of the Saccharomyces cerevisiae TEA/ATTS Family Transcription Factor Tec1p in Regulation of Gene Expression and Cellular Development , 2002, Eukaryotic Cell.

[83]  G. Fink,et al.  MAP kinase and cAMP filamentation signaling pathways converge on the unusually large promoter of the yeast FLO11 gene , 1999, The EMBO journal.