Piezo proteins are pore-forming subunits of mechanically activated channels

Abstract

Mechanotransduction has an important role in physiology. Biological processes including sensing touch and sound waves require as-yet-unidentified cation channels that detect pressure. Mouse Piezo1 (MmPiezo1) and MmPiezo2 (also called Fam38a and Fam38b, respectively) induce mechanically activated cationic currents in cells; however, it is unknown whether Piezo proteins are pore-forming ion channels or modulate ion channels. Here we show that Drosophila melanogaster Piezo (DmPiezo, also called CG8486) also induces mechanically activated currents in cells, but through channels with remarkably distinct pore properties including sensitivity to the pore blocker ruthenium red and single channel conductances. MmPiezo1 assembles as a 1.2-million-dalton homo-oligomer, with no evidence of other proteins in this complex. Purified MmPiezo1 reconstituted into asymmetric lipid bilayers and liposomes forms ruthenium-red-sensitive ion channels. These data demonstrate that Piezo proteins are an evolutionarily conserved ion channel family involved in mechanotransduction.

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Figure 1: Human cells expressing Drosophila Piezo (DmPiezo) show large mechanically activated currents.
Figure 2: Ruthenium red is a channel pore blocker of MmPiezo1- but not DmPiezo-induced currents.
Figure 3: MmPiezo1- and DmPiezo-induced stretch-activated channels have different conductances.
Figure 4: MmPiezo1 forms homo-oligomers.
Figure 5: MmPiezo1 forms ruthenium-red-sensitive ion channels.

Accession codes

Primary accessions

GenBank/EMBL/DDBJ

Data deposits

The DmPiezo sequence has been deposited in GenBank under accession number JQ425255.

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Acknowledgements

We thank M. H. Ulbrich for providing Ca2+ channel-, NMDA receptor- and CNG channel–GFP fusion constructs used as controls for photobleaching experiments. This research was supported by grants from the National Institutes of Dental and Craniofacial Research, Neurological Disorders, General Medical Sciences, and by The Genomics Institute of the Novartis Research Foundation. B.X. and J.G. are postdoctoral fellowship recipients from the American Heart Association and the NIH, respectively.

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B.C. performed and analysed electrophysiological experiments. B.X. performed and analysed biochemical experiments. J.S.S. and R.S. performed the reconstitution experiments and together with M.M. analysed the single channel data. J.G. and K.S.S. performed and analysed photo-bleaching experiments. S.E.K. cloned the Dmpiezo gene. M.S. initiated biochemical experiments. J.M. generated GFP-MmPiezo1 and the mRNA used for oocyte injection. A.E.D. provided technical help for oocyte experiments. A.P., B.C., B.X., J.G., J.S.S., R.S., and M.M. wrote the manuscript.

Corresponding authors

Correspondence to Mauricio Montal or Ardem Patapoutian.

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The authors declare no competing financial interests.

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Coste, B., Xiao, B., Santos, J. et al. Piezo proteins are pore-forming subunits of mechanically activated channels. Nature 483, 176–181 (2012). https://doi.org/10.1038/nature10812

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