Functional patchworking at the plasma membrane

Léon, Sébastien; Teis, David

doi:10.15252/embj.2018100144

Cited by 3 publications

(4 citation statements)

References 16 publications

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“…The protein samples were prepared and loaded onto a 5% stacking gel and 15% separating gel. After the samples were resolved by SDS-PAGE, they were transferred onto a 0.1 μm nitrocellulose transfer membrane (G161476, Whatman) under 3.5 mA/cm ( Léon and Teis, 2018 ) (E835, Consort) for 30 min at room temperature. After that, the proteins were probed with an HPR conjugated mouse anti-GFP antibody (working dilution: 1:5000) (SIGMA-ALDRICH).…”

Section: Methodsmentioning

confidence: 99%

“…It segments into coexisting compartments, which enable spatiotemporal segregation and coordination of different activities within the system (Zahumensky and Malinsky, 2019). For example, it has been recognized for a long time that there are heterogeneous lateral compartments in the plasma membrane of all cells (Léon and Teis, 2018). In fungi, plasma membrane lateral microdomains have been reported.…”

Section: Introductionmentioning

confidence: 99%

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The Composition and the Structure of MCC/Eisosomes in Neurospora crassa

Qin

Kempken

2020

Front. Microbiol.

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MCC/eisosomes are protein-organized domains in the plasma membrane of fungi and algae. However, the composition and function(s) of MCC/eisosomes in the filamentous fungus Neurospora crassa were previously unknown. To identify proteins that localize to MCC/eisosomes in N. crassa, we isolated proteins that co-purified with the core MCC/eisosome protein LSP-1, which was tagged with GFP. Proteins that co-fractionated with LSP-1:GFP were then identified by mass spectrometry. Eighteen proteins were GFP-tagged and used to identify six proteins that highly colocalized with the MCC/eisosome marker LSP-1:RFP, while five other proteins showed partial overlap with MCC/eisosomes. Seven of these proteins showed amino acid sequence homology with proteins known to localize to MCC/eisosomes in the yeast Saccharomyces cerevisiae. However, homologs of three proteins known to localize to MCC/eisosomes in S. cerevisiae (Can1, Pkh1/2, and Fhn1) were not found to colocalize with MCC/eisosome proteins in N. crassa by fluorescence microscopy. Interestingly, one new eisosome protein (glutamine-fructose-6-phosphate aminotransferase, gene ID: NCU07366) was detected in our studies. These findings demonstrate that there are interspecies differences of the protein composition of MCC/eisosomes. To gain further insight, molecular modeling and bioinformatics analysis of the identified proteins were used to propose the organization of MCC/eisosomes in N. crassa. A model will be discussed for how the broad range of functions predicted for the proteins localized to MCC/eisosomes, including cell wall synthesis, response and signaling, transmembrane transport, and actin organization, suggests that MCC/eisosomes act as organizing centers in the plasma membrane.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The Composition and the Structure of MCC/Eisosomes in Neurospora crassa

Qin

Kempken

2020

Front. Microbiol.

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“…Yeast cells uptake nutrients from their external environment through specific transporters that localise to the PM ( Jack et al, 2000 ; Léon and Teis, 2018 ). Regulation of these transporters at the PM allows for nutrient acquisition to be tightly controlled in response to cellular requirements.…”

Section: Introductionmentioning

confidence: 99%

The phosphatase Glc7 controls the eisosomal response to starvation via post-translational modification of Pil1

Paine

Laidlaw

Evans

et al. 2023

Journal of Cell Science

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The yeast plasma membrane (PM) is organised into specific subdomains that regulate surface membrane proteins. Surface transporters actively uptake nutrients in particular regions of the PM where they are also susceptible to substrate induced endocytosis. However, transporters also diffuse into distinct subdomains termed eisosomes, where they are protected from endocytosis. Although most nutrient transporter populations are downregulated in the vacuole following glucose starvation, a small pool is retained in eisosomes to provide efficient recovery from starvation. We find the core eisosome subunit Pil1, a Bin, Amphiphysin and Rvs (BAR) domain protein required for eisosome biogenesis, is phosphorylated primarily by the kinase Pkh2. In response to acute glucose starvation, Pil1 is rapidly dephosphorylated. Enzyme localisation and activity screens implicate the phosphatase Glc7 is the primary enzyme responsible for Pil1 dephosphorylation. Both depletion of GLC7 and phospho-ablative or phospho-mimetic mutations of Pil1 correlate with Pil1 phosphorylation status, failure to properly retain transporters in eisosomes, and results in defective starvation recovery. We propose precise posttranslational control of Pil1 modulates nutrient transporter retention within eisosomes depending on extracellular nutrient levels, to maximise recovery following starvation.

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“…The plasma membrane (PM) of eukaryotic cells is organised into distinct domains of specific lipids and proteins (Kraft, 2013). In the budding yeast Saccharomyces cerevisiae , microdomains of the PM have been characterised by fluorescence microscopy and shown to house distinct protein populations (Léon and Teis, 2018). Original localisation studies distinguished between the hexose transporter Hxt1, that is uniformly dispersed across the surface and other proteins that were found in discrete, non-overlapping regions (Malínská et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

The phosphatase Glc7 controls eisosomal response to starvation via posttranslational modification of Pil1

Paine

Evans

Laidlaw

et al. 2022

Preprint

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The yeast plasma membrane (PM) is organised into specific subdomains that can regulate the activity of surface membrane proteins localised within them. Nutrient transporters actively uptake substrates in particular regions of the PM where they are also susceptible to the endocytic machinery for substrate induced degradation. However, transporters also diffuse into distinct subdomains termed eisosomes, where they are inactive for substrate uptake and are protected from endocytosis. Although most nutrient transporter populations are sorted to the vacuole for degradation during glucose starvation, a small pool are retained in eisosomes. Sequestering this small pool of transporters during nutrient stress is essential for efficient recovery from starvation following a return to replete conditions. However, the mechanisms controlling this process at a biochemical level are poorly defined. We find the core eisosome subunit Pil1, a Bin, Amphiphysin and Rvs. (BAR) domain protein involved in membrane dynamics required for eisosome biogenesis, is primarily phosphorylated by Pkh2 but that Pil1 dephosphorylation occurs during acute glucose starvation. We screened for enzyme localisation and activity to implicate the essential phosphatase Glc7 as the primary enzyme responsible for Pil1 dephosphorylation. Manipulation of GLC7 expression correlates with Pil1 hypo/hyper phosphorylation. Furthermore, glc7 mutants and Pil1-phosphomutants are defective in recovering from glucose starvation. We propose precise posttranslational control of Pil1 modulates nutrient transporter retention within eisosomes depending on cellular nutritional status, to maximise recovery from starvation.

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Functional patchworking at the plasma membrane

Cited by 3 publications

References 16 publications

The Composition and the Structure of MCC/Eisosomes in Neurospora crassa

The Composition and the Structure of MCC/Eisosomes in Neurospora crassa

The phosphatase Glc7 controls the eisosomal response to starvation via post-translational modification of Pil1

The phosphatase Glc7 controls eisosomal response to starvation via posttranslational modification of Pil1

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