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PMID: 26785635 Published · epublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Removal of the mechanoprotective influence of the cytoskeleton reveals PIEZO1 is gated by bilayer tension.

Nature communications ·Vol. 7 ·2016-01-20 ·Pages 10366

Cox CD, Bae C, Ziegler L, Hartley S, Nikolova-Krstevski V, Rohde PR, Ng CA, Sachs F, Gottlieb PA, Martinac B

Abstract

Mechanosensitive ion channels are force-transducing enzymes that couple mechanical stimuli to ion flux. Understanding the gating mechanism of mechanosensitive channels is challenging because the stimulus seen by the channel reflects forces shared between the membrane, cytoskeleton and extracellular matrix. Here we examine whether the mechanosensitive channel PIEZO1 is activated by force-transmission through the bilayer. To achieve this, we generate HEK293 cell membrane blebs largely free of cytoskeleton. Using the bacterial channel MscL, we calibrate the bilayer tension demonstrating that activation of MscL in blebs is identical to that in reconstituted bilayers. Utilizing a novel PIEZO1-GFP fusion, we then show PIEZO1 is activated by bilayer tension in bleb membranes, gating at lower pressures indicative of removal of the cortical cytoskeleton and the mechanoprotection it provides. Thus, PIEZO1 channels must sense force directly transmitted through the bilayer.

MeSH Terms
Cell Survival/physiology Cytoskeleton/metabolism HEK293 Cells Humans Ion Channels/metabolism Lipid Bilayers/metabolism
Chemicals
Ion Channels Lipid Bilayers PIEZO1 protein, human
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Cox Charles D
Victor Chang Cardiac Research Institute, Darlinghurst, New South Wales 2010, Australia.
Bae Chilman
Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, New York 14214, USA.
Ziegler Lynn
Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, New York 14214, USA.
Hartley Silas
Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, New York 14214, USA.
Nikolova-Krstevski Vesna
Victor Chang Cardiac Research Institute, Darlinghurst, New South Wales 2010, Australia.
Rohde Paul R
Victor Chang Cardiac Research Institute, Darlinghurst, New South Wales 2010, Australia.
Ng Chai-Ann
Victor Chang Cardiac Research Institute, Darlinghurst, New South Wales 2010, Australia. | St Vincent's Clinical School, University of New South Wales, Darlinghurst, New South Wales 2010, Australia.
Sachs Frederick ORCID
Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, New York 14214, USA. | The Centre for Single Molecule Biophysics, State University of New York at Buffalo, Buffalo, New York 14214, USA.
Gottlieb Philip A
Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, New York 14214, USA. | The Centre for Single Molecule Biophysics, State University of New York at Buffalo, Buffalo, New York 14214, USA.
Martinac Boris ORCID
Victor Chang Cardiac Research Institute, Darlinghurst, New South Wales 2010, Australia. | St Vincent's Clinical School, University of New South Wales, Darlinghurst, New South Wales 2010, Australia.
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Article Info
Journal
Nature communications
Abbr.
Nat Commun
ISSN
2041-1723
DOI
10.1038/ncomms10366
Published
2016-01-20
Epub
2016-00-20
Pages
10366
Language
English
Region
England
NLM ID
101528555
PMCID
PMC4735864
Subset
IM
Grants
NHLBI NIH HHS · R01 HL054887 · United States
NHLBI NIH HHS · NIH 5R01HL054887 · United States
External Links
PubMed source DOI: 10.1038/ncomms10366 PMC Full Text View journal details
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