A steep phosphoinositide bis-phosphate gradient forms during fungal filamentous growth

Vernay, Aurélia; Schaub, Sébastien; Guillas, Isabelle; Bassilana, Martine; Arkowitz, Robert A.

doi:10.1083/jcb.201203099

Cited by 59 publications

(96 citation statements)

References 94 publications

(148 reference statements)

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“…The PI4P5K Mss4p is critical for filamentous growth in Candida albicans (Vernay et al, 2012) and Neurospora crassa (Seiler and Plamann, 2003;Mähs et al, 2012). In the pathogenic fungi C. albicans, Mss4p is critical for invasive filamentous growth and, in particular, the switch between the yeast and filamentous forms (Vernay et al, 2012). In N. crassa, mss4 mutants were identified in a screen for hyphal morphogenesis defects and exhibited general polarity defects (Seiler and Plamann, 2003).…”

Section: Discussionmentioning

confidence: 99%

“…5D). We used a tailor-made MATLAB program to analyze the signal concentration over long axis of the cell (Vernay et al, 2012) and Fig. 5E shows that in the wild-type strain there was an average 2-fold increase in PtdIns(4,5)P 2 concentration going from the back of the mother cell to the tip of the bud.…”

Section: Induction Of the Flo11p Cell Adhesion Flocculin Is Reduced Imentioning

confidence: 99%

“…We used the PtdIns(4,5)P 2 reporter that we had optimized for use in C. albicans (Vernay et al, 2012) to visualize plasma membrane PtdIns(4,5)P 2 in S. cerevisiae. This GFP-PH Plcd -PH Plcd -GFP reporter, driven by the GAL1 promoter to minimize toxicity associated with long-term expression was integrated at the URA3 locus.…”

Section: Induction Of the Flo11p Cell Adhesion Flocculin Is Reduced Imentioning

confidence: 99%

“…Images (3060.15 mm z-sections) were deconvolved using softWoRx software and maximum projections generated. All GFP and RFP images (except PtdIns(4,5)P 2 analyses) were acquired on an Andor Technology Revolution-XD spinning-disk laser confocal microscope (UplanSApo 1.4 NA 1006 objective) controlled by v2 iQ2 software with 488-nm or 561-nm laser excitation and 14061406400 nm voxel size (Vernay et al, 2012). Sum projections were generated by ImageJ.…”

Section: Microscopy and Image Analysesmentioning

confidence: 99%

“…To visualize plasma membrane PtdIns(4,5)P 2 a XhoI/NotI GFP-PH Plcd -PH Plcd -GFP fragment optimized for expression in C. albicans (Vernay et al, 2012) from pExpARG-pADH1GFP-PH Plcd -PH Plcd -GFP was cloned into pRS406 with a GAL1-10 promoter inserted at the XhoI site. This resulting plasmid, p406GALpGFP-PH Plcd -PH Plcd -GFP was linearized with StuI and transformed into wild-type (RAY1990) and mss4-f12 strains (RAY1991).…”

Section: Growth Conditions Strains and Plasmidsmentioning

confidence: 99%

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Phosphoinositide-bis-phosphate is required for Saccharomyces cerevisiae invasive growth

Guillas

Vernay

Vitagliano

et al. 2013

Journal of Cell Science

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SummaryPhosphatidylinositol phosphates are important regulators of processes such as the cytoskeleton organization, membrane trafficking and gene transcription, which are all crucial for polarized cell growth. In particular, phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P 2 ] has essential roles in polarized growth as well as in cellular responses to stress. In the yeast Saccharomyces cerevisiae, the sole phosphatidylinositol-4-phosphate 5-kinase (PI4P5K) Mss4p is essential for generating plasma membrane PtdIns(4,5)P 2 . Here, we show that Mss4p is required for yeast invasive growth in low-nutrient conditions. We isolated specific mss4 mutants that were defective in cell elongation, induction of the Flo11p flocculin, adhesion and cell wall integrity. We show that mss4-f12 cells have reduced plasma membrane PtdIns(4,5)P 2 levels as well as a defect in its polarized distribution, yet Mss4-f12p is catalytically active in vitro. In addition, the Mss4-f12 protein was defective in localizing to the plasma membrane. Furthermore, addition of cAMP, but not an activated MAPKKK allele, partially restored the invasive growth defect of mss4-f12 cells. Taken together, our results indicate that plasma membrane PtdIns(4,5)P 2 is crucial for yeast invasive growth and suggest that this phospholipid functions upstream of the cAMP-dependent protein kinase A signaling pathway.

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Section: Discussionmentioning

confidence: 99%

Section: Induction Of the Flo11p Cell Adhesion Flocculin Is Reduced Imentioning

confidence: 99%

Section: Induction Of the Flo11p Cell Adhesion Flocculin Is Reduced Imentioning

confidence: 99%

Section: Microscopy and Image Analysesmentioning

confidence: 99%

Section: Growth Conditions Strains and Plasmidsmentioning

confidence: 99%

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Phosphoinositide-bis-phosphate is required for Saccharomyces cerevisiae invasive growth

Guillas

Vernay

Vitagliano

et al. 2013

Journal of Cell Science

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Lipid Domains and Membrane (Re)Shaping: From Biophysics to Biology

Léonard

Alsteens

Dumitru

et al. 2017

Springer Series in Biophysics

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Regulation of yeast polarized exocytosis by phosphoinositide lipids

Volpiana,

Nenadic,

Beh

2024

Cell. Mol. Life Sci.

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Phosphoinositides help steer membrane trafficking routes within eukaryotic cells. In polarized exocytosis, which targets vesicular cargo to sites of polarized growth at the plasma membrane (PM), the two phosphoinositides phosphatidylinositol 4-phosphate (PI4P) and its derivative phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) pave the pathway for vesicle transport from the Golgi to the PM. PI4P is a critical regulator of mechanisms that shape late Golgi membranes for vesicle biogenesis and release. Although enriched in vesicle membranes, PI4P is inexplicably removed from post-Golgi vesicles during their transit to the PM, which drives subsequent steps in exocytosis. At the PM, PI(4,5)P2 recruits effectors that establish polarized membrane sites for targeting the vesicular delivery of secretory cargo. The budding yeast Saccharomyces cerevisiae provides an elegant model to unravel the complexities of phosphoinositide regulation during polarized exocytosis. Here, we review how PI4P and PI(4,5)P2 promote yeast vesicle biogenesis, exocyst complex assembly and vesicle docking at polarized cortical sites, and suggest how these steps might impact related mechanisms of human disease.

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A steep phosphoinositide bis-phosphate gradient forms during fungal filamentous growth

Abstract: A gradient of PI(4,5)P2 formed by phospholipid synthesis, diffusion, and regulated turnover is crucial for filamentous growth.

Cited by 59 publications

References 94 publications

Phosphoinositide-bis-phosphate is required for Saccharomyces cerevisiae invasive growth

Phosphoinositide-bis-phosphate is required for Saccharomyces cerevisiae invasive growth

Lipid Domains and Membrane (Re)Shaping: From Biophysics to Biology

Regulation of yeast polarized exocytosis by phosphoinositide lipids

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