Proteins involved in sterol synthesis interact with Ste20 and regulate cell polarity

Tiedje, Christopher; Holland, Daniel G.; Just, Ursula; Höfken, Thomas

doi:10.1242/jcs.009860

Cited by 48 publications

(67 citation statements)

References 77 publications

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“…The split-ubiquitin screen using STE20 as bait is described in Tiedje et al (2007). For the interaction assays, 10 5 ste20⌬ cells carrying the split-ubiquitin plasmids were spotted on SC-His/Leu and SC-His/Leu/Ura plates and were grown for 2 d at 30°C.…”

Section: Split-ubiquitin Techniquementioning

confidence: 99%

“…Previously, we described a screen to identify novel interactors of Ste20 by using the split-ubiquitin technique (Tiedje et al, 2007). This method is based on the ability of N-and C-terminal halves of ubiquitin to assemble into a quasinative ubiquitin (Johnsson and Varshavsky, 1994;Wittke et al, 1999).…”

Section: Paks Interact With the Transcriptional Regulator Sut1mentioning

confidence: 99%

“…Very little is known about Skm1, and no clear function has been attributed to this PAK (Martín et al, 1997). Interestingly, Ste20 and Cla4 interact with Erg4, Cbr1 and Ncp1, which are all involved in sterol biosynthesis, and the deletion of the corresponding genes results in various polarity defects, suggesting that sterol biosynThis article was published online ahead of print in MBC in Press (http://www.molbiolcell.org/cgi/doi/10.1091/mbc.E09 -01-0034) on September 30, 2009. thesis is crucial for cell polarization (Ni and Snyder, 2001;Tiedje et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

“…Budding yeast expresses three PAKs: Ste20, Cla4, and Skm1. To gain insight into molecular mechanisms of Ste20 action, we screened for interactors of Ste20 using the split-ubiquitin technique (Tiedje et al, 2007). Here, we show that Sut1, a transcriptional regulator that controls sterol uptake (Bourot and Karst, 1995;Ness et al, 2001), forms a complex with Ste20.…”

mentioning

confidence: 99%

“…© 2009 by The American Society for Cell Biology thesis is crucial for cell polarization (Ni and Snyder, 2001;Tiedje et al, 2007). Many aspects of sterol homeostasis are conserved between yeast and human, and ergosterol, the predominant sterol of yeast, is structurally and functionally related to sterols of higher eukaryotes (Sturley, 2000).…”

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confidence: 99%

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The Cdc42 Effectors Ste20, Cla4, and Skm1 Down-Regulate the Expression of Genes Involved in Sterol Uptake by a Mitogen-activated Protein Kinase-independent Pathway

Lin

Unden

Jacquier

et al. 2009

MBoC

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In Saccharomyces cerevisiae, the Rho-type GTPase Cdc42 regulates polarized growth through its effectors, including the p21-activated kinases (PAKs) Ste20, Cla4, and Skm1. Previously, we demonstrated that Ste20 interacts with several proteins involved in sterol synthesis that are crucial for cell polarization. Under anaerobic conditions, sterols cannot be synthesized and need to be imported into cells. Here, we show that Ste20, Cla4, and Skm1 form a complex with Sut1, a transcriptional regulator that promotes sterol uptake. All three PAKs can translocate into the nucleus and down-regulate the expression of genes involved in sterol uptake, including the Sut1 targets AUS1 and DAN1 by a novel mechanism. Consistently, deletion of either STE20, CLA4, or SKM1 results in an increased sterol influx and PAK overexpression inhibits sterol uptake. For Ste20, we demonstrate that the down-regulation of gene expression requires nuclear localization and kinase activity of Ste20. Furthermore, the Ste20-mediated control of expression of sterol uptake genes depends on SUT1 but is independent of a mitogen-activated protein kinase signaling cascade. Together, these observations suggest that PAKs translocate into the nucleus, where they modulate expression of sterol uptake genes via Sut1, thereby controlling sterol homeostasis. INTRODUCTIONThe small Rho GTPase Cdc42 plays a central role in the regulation of cellular polarity in eukaryotic cells (EtienneManneville, 2004;Jaffe and Hall, 2005). In the budding yeast Saccharomyces cerevisiae, Cdc42 regulates different types of polarized growth during various phases of its life cycle, including budding during vegetative growth, mating between haploid cells of opposite mating types, and filamentous growth upon nutrient limitation (Park and Bi, 2007). Cdc42 promotes polarized growth through multiple pathways, including polarization of the actin cytoskeleton and directed vesicle trafficking. Among the Cdc42 effectors that trigger these pathways are Ste20, Cla4 and Skm1, members of the p21-activated kinase (PAK) family of serine/threonine protein kinases (Hofmann et al., 2004;Park and Bi, 2007).Cdc42 recruits Ste20 and Cla4 from the cytoplasm and activates them at the plasma membrane at sites of polarized growth. Therefore, Ste20 and Cla4 are enriched at tips of buds and mating projections (Peter et al., 1996;Leberer et al., 1997;Holly and Blumer, 1999). The recruitment of Ste20 and Cla4 to these sites is governed by protein-protein as well as protein-lipid interactions. PAKs carry a conserved Cdc42/ Rac-interactive binding (CRIB) domain that mediates binding to Cdc42 and regulates their activity (Cvrckova et al., 1995;Peter et al., 1996;Leberer et al., 1997;Lamson et al., 2002;Ash et al., 2003). Ste20 and Cla4 also bind to Bem1, a scaffold protein that brings Cdc42, its activator Cdc24 and either Ste20 or Cla4, into proximity (Leeuw et al., 1998;Gulli et al., 2000;Bose et al., 2001;Yamaguchi et al., 2007). In addition, phosphoinositide-binding domains promote the association with membrane lip...

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Section: Split-ubiquitin Techniquementioning

confidence: 99%

Section: Paks Interact With the Transcriptional Regulator Sut1mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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The Cdc42 Effectors Ste20, Cla4, and Skm1 Down-Regulate the Expression of Genes Involved in Sterol Uptake by a Mitogen-activated Protein Kinase-independent Pathway

Lin

Unden

Jacquier

et al. 2009

MBoC

Self Cite

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Split‐Ubiquitin and the Split‐Protein Sensors: Chessman for the Endgame

Müller

Johnsson

2008

ChemBioChem

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The wide‐ranging phenotypes of ergosterol biosynthesis mutants, and implications for microbial cell factories

Johnston

Moses

Rosser

2020

Yeast

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Yeast strains have been used extensively as robust microbial cell factories for the production of bulk and fine chemicals, including biofuels (bioethanol), complex pharmaceuticals (antimalarial drug artemisinin and opioid pain killers), flavours, and fragrances (vanillin, nootkatone, and resveratrol). In many cases, it is of benefit to suppress or modify ergosterol biosynthesis during strain engineering, for example, to increase thermotolerance or to increase metabolic flux through an alternate pathway.However, the impact of modifying ergosterol biosynthesis on engineered strains is discussed sparsely in literature, and little attention has been paid to the implications of these modifications on the general health and well-being of yeast. Importantly, yeast with modified sterol content exhibit a wide range of phenotypes, including altered organization and dynamics of plasma membrane, altered susceptibility to chemical treatment, increased tolerance to high temperatures, and reduced tolerance to other stresses such as high ethanol, salt, and solute concentrations. Here, we review the wide-ranging phenotypes of viable Saccharomyces cerevisiae strains with altered sterol content and discuss the implications of these for yeast as microbial cell factories.

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Proteins involved in sterol synthesis interact with Ste20 and regulate cell polarity

Cited by 48 publications

References 77 publications

The Cdc42 Effectors Ste20, Cla4, and Skm1 Down-Regulate the Expression of Genes Involved in Sterol Uptake by a Mitogen-activated Protein Kinase-independent Pathway

The Cdc42 Effectors Ste20, Cla4, and Skm1 Down-Regulate the Expression of Genes Involved in Sterol Uptake by a Mitogen-activated Protein Kinase-independent Pathway

Split‐Ubiquitin and the Split‐Protein Sensors: Chessman for the Endgame

The wide‐ranging phenotypes of ergosterol biosynthesis mutants, and implications for microbial cell factories

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