Calcium-stimulated disassembly of focal adhesions mediated by an orp3/iqsec1 complex

Ryan S. D’souza; Jun Y. Lim; Alper Turgut; Kelly Servage; Junmei Zhang; Kim Orth; Nisha G. Sosale; Matthew J. Lazzara; Jeremy Allegood; James E. Casanova

doi:10.7554/eLife.54113

Calcium-stimulated disassembly of focal adhesions mediated by an orp3/iqsec1 complex

Ryan S. D’souza, Jun Y. Lim, Alper Turgut, Kelly Servage, Junmei Zhang, Kim Orth, Nisha G. Sosale, Matthew J. Lazzara, Jeremy Allegood, James E. Casanova

Research output: Contribution to journal › Article › peer-review

41 Scopus citations

Abstract

Coordinated assembly and disassembly of integrin-mediated focal adhesions (FAs) is essential for cell migration. Many studies have shown that FA disassembly requires Ca²⁺ influx, however our understanding of this process remains incomplete. Here, we show that Ca²⁺ influx via STIM1/Orai1 calcium channels, which cluster near FAs, leads to activation of the GTPase Arf5 via the Ca²⁺-activated GEF IQSec1, and that both IQSec1 and Arf5 activation are essential for adhesion disassembly. We further show that IQSec1 forms a complex with the lipid transfer protein ORP3, and that Ca²⁺ influx triggers PKC-dependent translocation of this complex to ER/plasma membrane (PM) contact sites adjacent to FAs. In addition to allosterically activating IQSec1, ORP3 also extracts PI4P from the PM, in exchange for phosphatidylcholine. ORP3-mediated lipid exchange is also important for FA turnover. Together, these findings identify a new pathway that links calcium influx to FA turnover during cell migration.

Original language	English (US)
Article number	e54113
Journal	eLife
Volume	9
DOIs	https://doi.org/10.7554/eLife.54113
State	Published - Apr 2020

ASJC Scopus subject areas

General Neuroscience
General Immunology and Microbiology
General Biochemistry, Genetics and Molecular Biology

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10.7554/eLife.54113

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@article{3afd4ac4b7cc4ce2a1253b99cf34fdab,

title = "Calcium-stimulated disassembly of focal adhesions mediated by an orp3/iqsec1 complex",

abstract = "Coordinated assembly and disassembly of integrin-mediated focal adhesions (FAs) is essential for cell migration. Many studies have shown that FA disassembly requires Ca2+ influx, however our understanding of this process remains incomplete. Here, we show that Ca2+ influx via STIM1/Orai1 calcium channels, which cluster near FAs, leads to activation of the GTPase Arf5 via the Ca2+-activated GEF IQSec1, and that both IQSec1 and Arf5 activation are essential for adhesion disassembly. We further show that IQSec1 forms a complex with the lipid transfer protein ORP3, and that Ca2+ influx triggers PKC-dependent translocation of this complex to ER/plasma membrane (PM) contact sites adjacent to FAs. In addition to allosterically activating IQSec1, ORP3 also extracts PI4P from the PM, in exchange for phosphatidylcholine. ORP3-mediated lipid exchange is also important for FA turnover. Together, these findings identify a new pathway that links calcium influx to FA turnover during cell migration.",

author = "D{\textquoteright}souza, {Ryan S.} and Lim, {Jun Y.} and Alper Turgut and Kelly Servage and Junmei Zhang and Kim Orth and Sosale, {Nisha G.} and Lazzara, {Matthew J.} and Jeremy Allegood and Casanova, {James E.}",

note = "Funding Information: The authors thank David Castle, Bettina Winkler and Adam Greene for critically reading the manuscript, and Lloyd McMahon for help with image analysis. The authors are also grateful for DNA plasmids provided by Vesa Olkonnen, Rick Horwitz, Olaf Idevall-Hagren, David Castle, Isabelle Derre, Tamas Balla, Pietro De Camilli, Andrew Thorburn and John Ngsee. This work was supported by NIH grant (1RO1GM127361) to JEC. Funding Information: The authors thank David Castle, Bettina Winkler and Adam Greene for critically reading the manuscript, and Lloyd McMahon for help with image analysis. The authors are also grateful for DNA plas-mids provided by Vesa Olkonnen, Rick Horwitz, Olaf Idevall-Hagren, David Castle, Isabelle Derre, Tamas Balla, Pietro De Camilli, Andrew Thorburn and John Ngsee. This work was supported by NIH grant (1RO1GM127361) to JEC. Publisher Copyright: Copyright D{\textquoteright}Souza et al.",

year = "2020",

month = apr,

doi = "10.7554/eLife.54113",

language = "English (US)",

volume = "9",

journal = "eLife",

issn = "2050-084X",

publisher = "eLife Sciences Publications",

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T1 - Calcium-stimulated disassembly of focal adhesions mediated by an orp3/iqsec1 complex

AU - D’souza, Ryan S.

AU - Lim, Jun Y.

AU - Turgut, Alper

AU - Servage, Kelly

AU - Zhang, Junmei

AU - Orth, Kim

AU - Sosale, Nisha G.

AU - Lazzara, Matthew J.

AU - Allegood, Jeremy

AU - Casanova, James E.

N1 - Funding Information: The authors thank David Castle, Bettina Winkler and Adam Greene for critically reading the manuscript, and Lloyd McMahon for help with image analysis. The authors are also grateful for DNA plasmids provided by Vesa Olkonnen, Rick Horwitz, Olaf Idevall-Hagren, David Castle, Isabelle Derre, Tamas Balla, Pietro De Camilli, Andrew Thorburn and John Ngsee. This work was supported by NIH grant (1RO1GM127361) to JEC. Funding Information: The authors thank David Castle, Bettina Winkler and Adam Greene for critically reading the manuscript, and Lloyd McMahon for help with image analysis. The authors are also grateful for DNA plas-mids provided by Vesa Olkonnen, Rick Horwitz, Olaf Idevall-Hagren, David Castle, Isabelle Derre, Tamas Balla, Pietro De Camilli, Andrew Thorburn and John Ngsee. This work was supported by NIH grant (1RO1GM127361) to JEC. Publisher Copyright: Copyright D’Souza et al.

PY - 2020/4

Y1 - 2020/4

N2 - Coordinated assembly and disassembly of integrin-mediated focal adhesions (FAs) is essential for cell migration. Many studies have shown that FA disassembly requires Ca2+ influx, however our understanding of this process remains incomplete. Here, we show that Ca2+ influx via STIM1/Orai1 calcium channels, which cluster near FAs, leads to activation of the GTPase Arf5 via the Ca2+-activated GEF IQSec1, and that both IQSec1 and Arf5 activation are essential for adhesion disassembly. We further show that IQSec1 forms a complex with the lipid transfer protein ORP3, and that Ca2+ influx triggers PKC-dependent translocation of this complex to ER/plasma membrane (PM) contact sites adjacent to FAs. In addition to allosterically activating IQSec1, ORP3 also extracts PI4P from the PM, in exchange for phosphatidylcholine. ORP3-mediated lipid exchange is also important for FA turnover. Together, these findings identify a new pathway that links calcium influx to FA turnover during cell migration.

AB - Coordinated assembly and disassembly of integrin-mediated focal adhesions (FAs) is essential for cell migration. Many studies have shown that FA disassembly requires Ca2+ influx, however our understanding of this process remains incomplete. Here, we show that Ca2+ influx via STIM1/Orai1 calcium channels, which cluster near FAs, leads to activation of the GTPase Arf5 via the Ca2+-activated GEF IQSec1, and that both IQSec1 and Arf5 activation are essential for adhesion disassembly. We further show that IQSec1 forms a complex with the lipid transfer protein ORP3, and that Ca2+ influx triggers PKC-dependent translocation of this complex to ER/plasma membrane (PM) contact sites adjacent to FAs. In addition to allosterically activating IQSec1, ORP3 also extracts PI4P from the PM, in exchange for phosphatidylcholine. ORP3-mediated lipid exchange is also important for FA turnover. Together, these findings identify a new pathway that links calcium influx to FA turnover during cell migration.

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JO - eLife

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ER -

Calcium-stimulated disassembly of focal adhesions mediated by an orp3/iqsec1 complex

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