Single-molecule atomic force microscopy reveals clustering of the yeast plasma-membrane sensor Wsc1.

Signalling is a key feature of living cells which frequently involves the local clustering of specific proteins in the plasma membrane. How such protein clustering is achieved within membrane microdomains ("rafts") is an important, yet largely unsolved problem in cell biology. The plasma m...

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Autores principales:	Jürgen J Heinisch, Vincent Dupres, Sabrina Wilk, Arne Jendretzki, Yves F Dufrêne
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2010
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/eec424fecb6d46b190c173c821b6d5d4
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spelling	oai:doaj.org-article:eec424fecb6d46b190c173c821b6d5d42021-12-02T20:20:56ZSingle-molecule atomic force microscopy reveals clustering of the yeast plasma-membrane sensor Wsc1.1932-620310.1371/journal.pone.0011104https://doaj.org/article/eec424fecb6d46b190c173c821b6d5d42010-06-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/20559440/?tool=EBIhttps://doaj.org/toc/1932-6203Signalling is a key feature of living cells which frequently involves the local clustering of specific proteins in the plasma membrane. How such protein clustering is achieved within membrane microdomains ("rafts") is an important, yet largely unsolved problem in cell biology. The plasma membrane of yeast cells represents a good model to address this issue, since it features protein domains that are sufficiently large and stable to be observed by fluorescence microscopy. Here, we demonstrate the ability of single-molecule atomic force microscopy to resolve lateral clustering of the cell integrity sensor Wsc1 in living Saccharomyces cerevisiae cells. We first localize individual wild-type sensors on the cell surface, revealing that they form clusters of approximately 200 nm size. Analyses of three different mutants indicate that the cysteine-rich domain of Wsc1 has a crucial, not yet anticipated function in sensor clustering and signalling. Clustering of Wsc1 is strongly enhanced in deionized water or at elevated temperature, suggesting its relevance in proper stress response. Using in vivo GFP-localization, we also find that non-clustering mutant sensors accumulate in the vacuole, indicating that clustering may prevent endocytosis and sensor turnover. This study represents the first in vivo single-molecule demonstration for clustering of a transmembrane protein in S. cerevisiae. Our findings indicate that in yeast, like in higher eukaryotes, signalling is coupled to the localized enrichment of sensors and receptors within membrane patches.Jürgen J HeinischVincent DupresSabrina WilkArne JendretzkiYves F DufrênePublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 5, Iss 6, p e11104 (2010)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jürgen J Heinisch Vincent Dupres Sabrina Wilk Arne Jendretzki Yves F Dufrêne Single-molecule atomic force microscopy reveals clustering of the yeast plasma-membrane sensor Wsc1.
description	Signalling is a key feature of living cells which frequently involves the local clustering of specific proteins in the plasma membrane. How such protein clustering is achieved within membrane microdomains ("rafts") is an important, yet largely unsolved problem in cell biology. The plasma membrane of yeast cells represents a good model to address this issue, since it features protein domains that are sufficiently large and stable to be observed by fluorescence microscopy. Here, we demonstrate the ability of single-molecule atomic force microscopy to resolve lateral clustering of the cell integrity sensor Wsc1 in living Saccharomyces cerevisiae cells. We first localize individual wild-type sensors on the cell surface, revealing that they form clusters of approximately 200 nm size. Analyses of three different mutants indicate that the cysteine-rich domain of Wsc1 has a crucial, not yet anticipated function in sensor clustering and signalling. Clustering of Wsc1 is strongly enhanced in deionized water or at elevated temperature, suggesting its relevance in proper stress response. Using in vivo GFP-localization, we also find that non-clustering mutant sensors accumulate in the vacuole, indicating that clustering may prevent endocytosis and sensor turnover. This study represents the first in vivo single-molecule demonstration for clustering of a transmembrane protein in S. cerevisiae. Our findings indicate that in yeast, like in higher eukaryotes, signalling is coupled to the localized enrichment of sensors and receptors within membrane patches.
format	article
author	Jürgen J Heinisch Vincent Dupres Sabrina Wilk Arne Jendretzki Yves F Dufrêne
author_facet	Jürgen J Heinisch Vincent Dupres Sabrina Wilk Arne Jendretzki Yves F Dufrêne
author_sort	Jürgen J Heinisch
title	Single-molecule atomic force microscopy reveals clustering of the yeast plasma-membrane sensor Wsc1.
title_short	Single-molecule atomic force microscopy reveals clustering of the yeast plasma-membrane sensor Wsc1.
title_full	Single-molecule atomic force microscopy reveals clustering of the yeast plasma-membrane sensor Wsc1.
title_fullStr	Single-molecule atomic force microscopy reveals clustering of the yeast plasma-membrane sensor Wsc1.
title_full_unstemmed	Single-molecule atomic force microscopy reveals clustering of the yeast plasma-membrane sensor Wsc1.
title_sort	single-molecule atomic force microscopy reveals clustering of the yeast plasma-membrane sensor wsc1.
publisher	Public Library of Science (PLoS)
publishDate	2010
url	https://doaj.org/article/eec424fecb6d46b190c173c821b6d5d4
work_keys_str_mv	AT jurgenjheinisch singlemoleculeatomicforcemicroscopyrevealsclusteringoftheyeastplasmamembranesensorwsc1 AT vincentdupres singlemoleculeatomicforcemicroscopyrevealsclusteringoftheyeastplasmamembranesensorwsc1 AT sabrinawilk singlemoleculeatomicforcemicroscopyrevealsclusteringoftheyeastplasmamembranesensorwsc1 AT arnejendretzki singlemoleculeatomicforcemicroscopyrevealsclusteringoftheyeastplasmamembranesensorwsc1 AT yvesfdufrene singlemoleculeatomicforcemicroscopyrevealsclusteringoftheyeastplasmamembranesensorwsc1
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Single-molecule atomic force microscopy reveals clustering of the yeast plasma-membrane sensor Wsc1.

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