Differential regulation of Tec1 by Fus3 and Kss1 confers signaling specificity in yeast development

Brückner, Stefan; Köhler, Tim; Braus, Gerhard H.; Heise, Barbara; Bolte, Melanie; Mösch, Hans‐Ulrich

doi:10.1007/s00294-004-0545-1

Cited by 52 publications

(77 citation statements)

References 46 publications

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“…Ubiquitin attachment typically leads to rapid capture and degradation of the substrate by the proteasome protease complex (27), and it was recently documented that Tec1 undergoes phosphorylationdependent ubiquitination and accelerated degradation in pheromonestimulated cells (42)(43)(44). Moreover, modification by SUMO and ubiquitin are often antagonistic and can sometimes occur on the same Lys residue (45,46).…”

Section: Resultsmentioning

confidence: 99%

Pheromone-regulated Sumoylation of Transcription Factors That Mediate the Invasive to Mating Developmental Switch in Yeast

Wang

Dohlman

2006

Journal of Biological Chemistry

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A fundamental question in biology is how different signaling pathways use common signaling proteins to attain different developmental outcomes. The yeast transcription factor Ste12 is required in at least two distinct signaling processes, each regulated by many of the same protein kinases. Whereas Ste12-Ste12 homodimers promote transcription of genes required for mating, Ste12-Tec1 heterodimers activate genes required for invasive growth. We report that Ste12 and Tec1 undergo covalent modification by the ubiquitin-related modifier SUMO. Stimulation by mating pheromone promotes sumoylation of Ste12 and diminishes the sumoylation of Tec1. In the absence of sumoylation Tec1 is more rapidly degraded. We propose that pheromone-regulated sumoylation of Ste12 and Tec1 promotes a developmental switch from the invasive to the mating differentiation program.The budding yeast Saccharomyces cerevisiae can initiate distinct developmental programs depending on the presence or absence of specific external cues. Mating is initiated when a and ␣ haploid cell types secrete and respond to cell type-specific pheromones acting through G protein-coupled receptors; once activated, the a and ␣ cells fuse to form an a/␣ diploid cell. Invasive or filamentous growth occurs in nutrientpoor conditions and is manifested by altered budding and formation of long branching filaments, as well as increased adherence and invasion of the substratum. Both developmental outcomes require activation of a protein kinase cascade comprised of Ste20, Ste11, Ste7, and Fus3 or Kss1 (1, 2).Fus3 and Kss1 are mitogen-activated protein (MAP) 2 kinases that phosphorylate substrates required for signaling in both the mating and invasive growth pathways. Phosphorylation of Ste12 promotes binding to a specific DNA sequence called the pheromone-response element (PRE) (3-6), where it initiates transcription of genes required for efficient mating (7-9). Ste12 can also assemble with another transcription factor, Tec1 (10 -13). Ste12-Tec1 heterodimers bind cooperatively to a distinct DNA sequence called the filamentation-and invasive-response element (FRE) (10, 11) present in the promoter region of genes that regulate invasive or pseudohyphal growth (9, 14). The kinase activity of Kss1 increases filamentation, whereas the kinase activity of Fus3 suppresses filamentation (1,8,10,15,16). Both pathways are negatively regulated by Ste12-binding proteins Dig1/Rst1 and Dig2/Rst2 (7,(17)(18)(19) as well as by Ste12 binding to the unphosphorylated and inactive form of Kss1 (20).Although the mechanisms of MAP kinase and transcription factor activation are well established, less understood is how signaling pathways that share the same components attain different developmental fates (21). Signal identity has been ascribed to differences in signal magnitude, duration, and frequency (22), as well as to the scaffolded association of protein kinase components (23). Even where such differences have been documented, the kinase signals must still be interpreted by nuclear transcription fa...

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Section: Resultsmentioning

confidence: 99%

Pheromone-regulated Sumoylation of Transcription Factors That Mediate the Invasive to Mating Developmental Switch in Yeast

Wang

Dohlman

2006

Journal of Biological Chemistry

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“…Phosphorylation of Fus3 and Kss1 was detected using a Phospho-p44/42 MAP kinase (Thr202/Tyr204) antibody from Cell Signaling Technol- (10). RNA was separated on a 1.4% agarose gel containing 3% formaldehyde and transferred onto nylon membrane as described previously (6). CDC42 and ACT1 transcripts were detected using gene-specific 32 P-labeled DNA probes.…”

Section: Methodsmentioning

confidence: 99%

“…Preparation of total cell extracts and subsequent Western blot analysis were performed as described previously (6). After transfer to a nitrocellulose membrane, proteins were detected using the enhanced chemiluminescence system and incubation with primary antibodies against Cdc42 or Fus3 (Santa Cruz Biotechnology, Santa Cruz, California), Cdc28 (kind gift from S. Irniger), or the myc epitope followed by incubation with a peroxidase-coupled secondary antibody (Santa Cruz Biotechnology, Santa Cruz, California).…”

Section: Methodsmentioning

confidence: 99%

“…Yeast culture medium was prepared as described previously (18). Invasive growth tests were described earlier (6).…”

Section: Methodsmentioning

confidence: 99%

“…Strains carrying FUS1-lacZ reporter constructs were grown to the exponential growth phase, and extracts were prepared and assayed for ␤-galactosidase activity as described previously (6). ␤-Galactosidase activity was normalized to the total protein in each extract with the following formula: (optical density at 420 nm ϫ 1.7)/(0.0045 ϫ protein concentration ϫ extract volume ϫ time).…”

Section: Methodsmentioning

confidence: 99%

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Role of Cdc42-Cla4 Interaction in the Pheromone Response of Saccharomyces cerevisiae

2007

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In Saccharomyces cerevisiae, the highly conserved Rho-type GTPase Cdc42 is essential for cell division and controls cellular development during mating and invasive growth. The role of Cdc42 in mating has been controversial, but a number of previous studies suggest that the GTPase controls the mitogen-activated protein (MAP) kinase cascade by activating the p21-activated protein kinase (PAK) Ste20. To further explore the role of Cdc42 in pheromone-stimulated signaling, we isolated novel alleles of CDC42 that confer resistance to pheromone. We find that in CDC42(V36A) and CDC42(V36A, I182T) mutant strains, the inability to undergo pheromone-induced cell cycle arrest correlates with reduced phosphorylation of the mating MAP kinases Fus3 and Kss1 and with a decrease in mating efficiency. Furthermore, Cdc42(V36A) and Cdc42(V36A, I182T) proteins show reduced interaction with the PAK Cla4 but not with Ste20. We also show that deletion of CLA4 in a CDC42(V36A, I182T) mutant strain suppresses pheromone resistance and that overexpression of CLA4 interferes with pheromone-induced cell cycle arrest and MAP kinase phosphorylation in CDC42 wild-type strains. Our data indicate that Cla4 has the potential to act as a negative regulator of the mating pathway and that this function of the PAK might be under control of Cdc42. In conclusion, our study suggests that control of pheromone signaling by Cdc42 not only depends on Ste20 but also involves interaction of the GTPase with Cla4.

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Scaffold proteins in MAP kinase signaling: more than simple passive activating platforms

Dard

Peter

2006

Bioessays

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Due to the central position of scaffold proteins in numerous signaling networks, especially in MAPK pathways, considerable efforts have been made to identify new scaffolds and to characterize their function and regulation. Most of our knowledge stems from studies of yeast MAPK scaffolds, but the identification of such scaffolds in higher eukaryotes provided a new dimension to this field and led to exciting and promising new insights into the regulation of MAPK signaling. In this review, we shortly summarize the well-established basic functions of scaffolds in yeast and highlight concepts emerging from recent studies in yeast and higher eukaryotes. In particular, we discuss how scaffolds may actively influence MAPK signaling by inducing conformational changes of bound kinases or substrates, by controlling the localization of activated MAPK and the extent and output of MAPK activation, and by modulating MAPK kinetics through the recruitment of phosphatases or ubiquitin-ligases. Finally, we summarize the current knowledge of scaffold regulation, and how these events may be functionally important for MAPK signaling.

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Differential regulation of Tec1 by Fus3 and Kss1 confers signaling specificity in yeast development

Cited by 52 publications

References 46 publications

Pheromone-regulated Sumoylation of Transcription Factors That Mediate the Invasive to Mating Developmental Switch in Yeast

Pheromone-regulated Sumoylation of Transcription Factors That Mediate the Invasive to Mating Developmental Switch in Yeast

Role of Cdc42-Cla4 Interaction in the Pheromone Response of Saccharomyces cerevisiae

Scaffold proteins in MAP kinase signaling: more than simple passive activating platforms

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