Flippase-mediated phospholipid asymmetry promotes fast Cdc42 recycling in dynamic maintenance of cell polarity

Das, Arupratan; Slaughter, Brian D.; Unruh, Jay R.; Bradford, W. David; Richard, Alexander; Rubinstein, Boris; Li, Rong

doi:10.1038/ncb2444

Cited by 99 publications

(109 citation statements)

References 42 publications

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“…A subsequent study obtained a better-constrained value for the abundance of Cdc42 (1 mM) [30], which we have adopted in the current model. In addition, we manually tuned some of the other parameters so that the model would reproduce three features extracted from imaging data: (i) the measured shape of the Cdc42 peak ( peak width at half height, 1.9 mm [31]); (ii) Cdc42 dynamics in the peak (FRAP recovery half-time, 3.5 s [32]) and our estimate of the amount of Cdc42 in the peak ( proportion of the total Cdc42, 4.6%).…”

Section: (I) Background On Model Developmentmentioning

confidence: 99%

Interaction between bud-site selection and polarity-establishment machineries in budding yeast

Savage

Lew

2013

Phil. Trans. R. Soc. B

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Saccharomyces cerevisiae yeast cells polarize in order to form a single bud in each cell cycle. Distinct patterns of bud-site selection are observed in haploid and diploid cells. Genetic approaches have identified the molecular machinery responsible for positioning the bud site: during bud formation, specific locations are marked with immobile landmark proteins. In the next cell cycle, landmarks act through the Ras-family GTPase Rsr1 to promote local activation of the conserved Rho-family GTPase, Cdc42. Additional Cdc42 accumulates by positive feedback, creating a concentrated patch of GTP-Cdc42, which polarizes the cytoskeleton to promote bud emergence. Using time-lapse imaging and mathematical modelling, we examined the process of bud-site establishment. Imaging reveals unexpected effects of the bud-site-selection system on the dynamics of polarity establishment, raising new questions about how that system may operate. We found that polarity factors sometimes accumulate at more than one site among the landmark-specified locations, and we suggest that competition between clusters of polarity factors determines the final location of the Cdc42 cluster. Modelling indicated that temporally constant landmark-localized Rsr1 would weaken or block competition, yielding more than one polarity site. Instead, we suggest that polarity factors recruit Rsr1, effectively sequestering it from other locations and thereby terminating landmark activity.

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Section: (I) Background On Model Developmentmentioning

confidence: 99%

Interaction between bud-site selection and polarity-establishment machineries in budding yeast

Savage

Lew

2013

Phil. Trans. R. Soc. B

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“…Display rate, 1 frame /12 s. Related to Figure 5C. ; b (Berepiki et al 2011); c (Dagdas et al 2012); d (Chen and Thorner 2007); e (Semighini and Harris 2008); f (Jones and Bennett 2011); g ; h (Artiles et al 2009); i (Das et al 2013); j (Rank and Rayment 2013); k (Krappmann et al 2007); l (Pulver et al 2013); m (Park et al 1997); n (Brace et al 2011); o (Knaus et al 2007); p (Berepiki and Read 2013) (Glass and Lee 1992); g (Ferreira et al 1998); h (Hammond et al 2012); i (Li et al 2005); j (Park et al 2011) …”

Section: Figure S2mentioning

confidence: 99%

New Insights Into the Roles of NADPH Oxidases in Sexual Development and Ascospore Germination in Sordaria macrospora

et al. 2014

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NADPH oxidase (NOX)-derived reactive oxygen species (ROS) act as signaling determinants that induce different cellular processes. To characterize NOX function during fungal development, we utilized the genetically tractable ascomycete Sordaria macrospora. Genome sequencing of a sterile mutant led us to identify the NADPH oxidase encoding nox1 as a gene required for fruiting body formation, regular hyphal growth, and hyphal fusion. These phenotypes are shared by Δnor1, lacking the NOX regulator NOR1. Further phenotypic analyses revealed a high correlation between increased ROS production and hyphal fusion deficiencies in Δnox1 and other sterile mutants. A genome-wide transcriptional profiling analysis of mycelia and isolated protoperithecia from wild type and Δnox1 revealed that nox1 inactivation affects the expression of genes related to cytoskeleton remodeling, hyphal fusion, metabolism, and mitochondrial respiration. Genetic analysis of Δnox2, lacking the NADPH oxidase 2 gene, Δnor1, and transcription factor deletion mutant Δste12, revealed a strict melanin-dependent ascospore germination defect, indicating a common genetic pathway for these three genes. We report that gsa3, encoding a G-protein a-subunit, and sac1, encoding cAMP-generating adenylate cyclase, act in a separate pathway during the germination process. The finding that cAMP inhibits ascospore germination in a melanin-dependent manner supports a model in which cAMP inhibits NOX2 activity, thus suggesting a link between both pathways. Our results expand the current knowledge on the role of NOX enzymes in fungal development and provide a frame to define upstream and downstream components of the NOX signaling pathways in fungi. D URING sexual reproduction, filamentous fungi generate complex fruiting bodies that contain and protect meiosporangia. We used the ascomycetous model fungus Sordaria macrospora to identify genes directly involved in fruiting body development (Kück et al. 2009;Engh et al. 2010;Kück et al. 2009). Due to its homothallic life style, S. macrospora is able to complete the sexual life cycle without the mating of strains with opposite sex, and therefore, fruiting body-deficient mutants can be recognized directly without the need for crossing experiments. In earlier work, we generated sterile mutants showing a developmental block after formation of young fruiting bodies (protoperithecia), but being unable to generate mature perithecia, and referred to these mutants as pro. Recently, we have applied next-generation genome re-sequencing to identify the genes affected in some of these mutants . Based on this approach, we now have characterized mutant pro32 and show that it carries a mutation in the nox1 gene encoding NAPDH oxidase 1 (NOX1).NADPH oxidase (NOX) enzymes are transmembrane proteins that are highly conserved among eukaryotes and produce reactive oxygen species (ROS) through the oxidation of NADPH (Lambeth 2004;Kawahara and Lambeth 2007). ROS have long been recognized as damaging agents due to uncontrolled oxidizing re...

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“…Functional studies revealed that flippases are involved in various vesicle transport pathways (5), and flippases in these functions are implicated in vesicle formation by inducing local membrane curvature (16,17). Flippases also regulate functions of membrane proteins by changing transbilayer phospholipid composition; changes in PE and PS content in the cytoplasmic leaflet of the plasma membrane regulate Cdc42p localization in polarized cell growth (18,19). However, other functions of flippases need to be investigated.…”

mentioning

confidence: 99%

Phospholipid Flippases Lem3p-Dnf1p and Lem3p-Dnf2p Are Involved in the Sorting of the Tryptophan Permease Tat2p in Yeast

Hachiro

Yamamoto

Nakano

et al. 2013

Journal of Biological Chemistry

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Background: Lem3p-Dnf1p and Lem3p-Dnf2p are phospholipid flippases that generate phospholipid asymmetry in yeast. Results:The tryptophan permease Tat2p is missorted from the trans-Golgi network to the vacuole in the lem3⌬ mutant. Conclusion: Phospholipid asymmetry supports plasma membrane transport of Tat2p by inhibiting its improper ubiquitination at the trans-Golgi network. Significance: Phospholipid asymmetry may be involved in proper sorting of membrane proteins.

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Flippase-mediated phospholipid asymmetry promotes fast Cdc42 recycling in dynamic maintenance of cell polarity

Cited by 99 publications

References 42 publications

Interaction between bud-site selection and polarity-establishment machineries in budding yeast

Interaction between bud-site selection and polarity-establishment machineries in budding yeast

New Insights Into the Roles of NADPH Oxidases in Sexual Development and Ascospore Germination in Sordaria macrospora

Phospholipid Flippases Lem3p-Dnf1p and Lem3p-Dnf2p Are Involved in the Sorting of the Tryptophan Permease Tat2p in Yeast

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