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The yeast Wsc1 cell surface sensor behaves like a nanospring in vivo

Nature Chemical Biology volume 5, pages 857–862 (2009)Cite this article

Abstract

Here we report on in vivo measurement of the mechanical behavior of a cell surface sensor using single-molecule atomic force microscopy. We focus on the yeast wall stress component sensor Wsc1, a plasma membrane protein that is thought to function as a rigid probe of the cell wall status. We first map the distribution of individual histidine-tagged sensors on living yeast cells by scanning the cell surface with atomic force microscopy tips carrying nitrilotriacetate groups. We then show that Wsc1 behaves like a linear nanospring that is capable of resisting high mechanical force and of responding to cell surface stress. Both a genomic pmt4 deletion and the insertion of a stretch of glycines in Wsc1 result in substantial alterations in protein spring properties, supporting the important role of glycosylation at the extracellular serine/threonine-rich region.

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Figure 1: AFM detects single Wsc1 sensors on live cells.
Figure 2: Wsc1 behaves as a linear spring.
Figure 3: The spring properties of Wsc1 require glycosylation of the serine/threonine region.
Figure 4: The nanospring behavior measured for elongated Wsc1 is not substantially altered by the Mid2 elongation.

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Acknowledgements

This work was supported by the Belgian National Foundation for Scientific Research (FNRS), the Université catholique de Louvain (Fonds Spéciaux de Recherche), the Région wallonne, the Federal Office for Scientific, Technical and Cultural Affairs (Interuniversity Poles of Attraction Programme), and the Research Department of the Communauté française de Belgique (Concerted Research Action). Work at the University of Osnabrück was funded by the Deutsche Forschungsgemeinschaft within the framework of the SFB431.

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Author notes

  1. Vincent Dupres and David Alsteens: These authors contributed equally to this work.

Authors and Affiliations

  1. Unité de Chimie des Interfaces, Université Catholique de Louvain, Louvain-la-Neuve, Belgium

    Vincent Dupres, David Alsteens & Yves F Dufrêne

  2. Universität Osnabrück, Fachbereich Biologie/Chemie, AG Genetik, Osnabrück, Germany

    Sabrina Wilk, Benjamin Hansen & Jürgen J Heinisch

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  1. Vincent Dupres
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Contributions

V.D., D.A., S.W., B.H., J.J.H. and Y.F.D. designed the experiments, analyzed the data and wrote the article. S.W., B.H. and J.J.H. performed the genetic manipulations. V.D. and D.A. collected the AFM data.

Corresponding authors

Correspondence to Jürgen J Heinisch or Yves F Dufrêne.

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Dupres, V., Alsteens, D., Wilk, S. et al. The yeast Wsc1 cell surface sensor behaves like a nanospring in vivo. Nat Chem Biol 5, 857–862 (2009). https://doi.org/10.1038/nchembio.220

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