Is there anyone out there?-Single-molecule atomic force microscopy meets yeast genetics to study sensor functions

The ability to react to environmental stress is a key feature of microbial cells, which frequently involves the fortification of their cell wall as a primary step. In the model yeast Saccharomyces cerevisiae the biosynthesis of the cell wall is regulated by the so-called cell wall integrity signal transduction pathway, which starts with the detection of cell surface stress by a small family of five membrane-spanning sensors (Wsc1-Wsc3, Mid2, Mtl1). Although genetic evidence indicated that these proteins act as mechanosensors, direct in vivo evidence for their function remained scarce. Here, we review a new approach integrating the tools and concepts of genetics with those of nanotechnology. We show how atomic force microscopy can be combined with advanced protein design by yeast genetics, to study the function and the mechanical properties of yeast sensors in living cells down to the single molecule level. We anticipate that this novel integrated technology will enable a paradigm shift in cell biology, so that pertinent questions can be addressed, such as the nanomechanics of single sensors and receptors, and how they distribute across the cell surface when they respond to extracellular stress.