Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 107, Issue 48, Pages 20744-20749Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1013893107
Keywords
single-molecule techniques; microbial adhesion; glycoproteins; fungi
Categories
Funding
- Fonds National de la Recherche Scientifique
- Universite Catholique de Louvain (Fonds Speciaux de Recherche)
- Region Wallonne
- Federal Office for Scientific, Technical and Cultural Affairs
- Research Department of the Communaute Francaise de Belgique (Concerted Research Action)
- National Institutes of Health [SC1 GM083756]
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Understanding how cell adhesion proteins form adhesion domains is a key challenge in cell biology. Here, we use single-molecule atomic force microscopy (AFM) to demonstrate the force-induced formation and propagation of adhesion nanodomains in living fungal cells, focusing on the covalently anchored cell-wall protein Als5p from Candida albicans. We show that pulling on single adhesins with AFM tips terminated with specific antibodies triggers the formation of adhesion domains of 100-500 nm and that the force-induced nanodomains propagate over the entire cell surface. Control experiments (with cells lacking Als5p, single-site mutation in the protein, bare tips, and tips modified with irrelevant antibodies) demonstrate that Als5p nanodomains result from protein redistribution triggered by force-induced conformational changes in the initially probed proteins, rather than from nonspecific cell-wall perturbations. Als5p remodeling is independent of cellular metabolic activity because heat-killed cells show the same behavior as live cells. Using AFM and fluorescence microscopy, we also find that nanodomains are formed within similar to 30 min and migrate at a speed of similar to 20 nm.min(-1), indicating that domain formation and propagation are slow, time-dependent processes. These results demonstrate that mechanical stimuli can trigger adhesion nanodomains in fungal cells and suggest that the force-induced clustering of adhesins may be a mechanism for activating cell adhesion.
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