Cluster nanoarchitecture and structural diversity of PIEZO1 in intact cells

The mechanically activated ion channel PIEZO1 confers mechanosensitivity to many cell types and is thus essential for numerous physiological processes triggered by mechanical cues ^1–4 . PIEZO1 channels assemble as propeller-shaped trimers ^5–7 , which supposedly undergo a transition from a curved conformational state into a flattened state upon mechanical activation ^8–11 . Whether this gating model also applies to PIEZO1 clusters and to what extent membrane curvature alters PIEZO1 conformation in living cells, is, however, still unclear. Here, we used MINFLUX nanoscopy to resolve the architecture of PIEZO1 clusters and demonstrate that PIEZO1 channels collectively deform the cell membrane into pit-shaped invaginations, which accommodate multiple channels and flatten in response to mechanical stimulation. Moreover, our data suggests that the deformability of the membrane and thus the conformation and sensitivity of PIEZO1 are altered by the architecture and the rigidity of the cytoskeleton. Together, our data provide previously unrecognized structural details about the subcellular distribution and the conformational variability of PIEZO1 in different cellular compartments, which challenge previously proposed hypotheses derived from studies of isolated channels and thus highlight the importance of examining PIEZO1 in its native environment, where additional factors contribute to channel function and structure.