Abstract
The conditional use of actin during clathrin-mediated endocytosis in mammalian cells suggests that the cell controls whether and how actin is used. Using a combination of biochemical reconstitution and mammalian cell culture, we elucidate a mechanism by which the coincidence of PI(4,5)P2 and PI(3)P in a curved vesicle triggers actin polymerization. At clathrin-coated pits, PI(3)P is produced by the INPP4A hydrolysis of PI(3,4)P2, and this is necessary for actin-driven endocytosis. Both Cdc42·guanosine triphosphate and SNX9 activate N-WASP-WIP- and Arp2/3-mediated actin nucleation. Membrane curvature, PI(4,5)P2, and PI(3)P signals are needed for SNX9 assembly via its PX-BAR domain, whereas signaling through Cdc42 is activated by PI(4,5)P2 alone. INPP4A activity is stimulated by high membrane curvature and synergizes with SNX9 BAR domain binding in a process we call curvature cascade amplification. We show that the SNX9-driven actin comets that arise on human disease-associated oculocerebrorenal syndrome of Lowe (OCRL) deficiencies are reduced by inhibiting PI(3)P production, suggesting PI(3)P kinase inhibitors as a therapeutic strategy in Lowe syndrome.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 3745-3765 |
| Number of pages | 21 |
| Journal | Journal of Cell Biology |
| Volume | 216 |
| Issue number | 11 |
| DOIs | |
| State | Published - Nov 1 2017 |
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ASJC Scopus subject areas
- Cell Biology
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Control of actin polymerization via the coincidence of phosphoinositides and high membrane curvature. / Daste, Frederic; Walrant, Astrid; Holst, Mikkel R.; Gadsby, Jonathan R.; Mason, Julia; Lee, Ji Eun; Brook, Daniel; Mettlen, Marcel; Larsson, Elin; Lee, Steven F.; Lundmark, Richard; Gallop, Jennifer L.
In: Journal of Cell Biology, Vol. 216, No. 11, 01.11.2017, p. 3745-3765.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Control of actin polymerization via the coincidence of phosphoinositides and high membrane curvature
AU - Daste, Frederic
AU - Walrant, Astrid
AU - Holst, Mikkel R.
AU - Gadsby, Jonathan R.
AU - Mason, Julia
AU - Lee, Ji Eun
AU - Brook, Daniel
AU - Mettlen, Marcel
AU - Larsson, Elin
AU - Lee, Steven F.
AU - Lundmark, Richard
AU - Gallop, Jennifer L.
PY - 2017/11/1
Y1 - 2017/11/1
N2 - The conditional use of actin during clathrin-mediated endocytosis in mammalian cells suggests that the cell controls whether and how actin is used. Using a combination of biochemical reconstitution and mammalian cell culture, we elucidate a mechanism by which the coincidence of PI(4,5)P2 and PI(3)P in a curved vesicle triggers actin polymerization. At clathrin-coated pits, PI(3)P is produced by the INPP4A hydrolysis of PI(3,4)P2, and this is necessary for actin-driven endocytosis. Both Cdc42·guanosine triphosphate and SNX9 activate N-WASP-WIP- and Arp2/3-mediated actin nucleation. Membrane curvature, PI(4,5)P2, and PI(3)P signals are needed for SNX9 assembly via its PX-BAR domain, whereas signaling through Cdc42 is activated by PI(4,5)P2 alone. INPP4A activity is stimulated by high membrane curvature and synergizes with SNX9 BAR domain binding in a process we call curvature cascade amplification. We show that the SNX9-driven actin comets that arise on human disease-associated oculocerebrorenal syndrome of Lowe (OCRL) deficiencies are reduced by inhibiting PI(3)P production, suggesting PI(3)P kinase inhibitors as a therapeutic strategy in Lowe syndrome.
AB - The conditional use of actin during clathrin-mediated endocytosis in mammalian cells suggests that the cell controls whether and how actin is used. Using a combination of biochemical reconstitution and mammalian cell culture, we elucidate a mechanism by which the coincidence of PI(4,5)P2 and PI(3)P in a curved vesicle triggers actin polymerization. At clathrin-coated pits, PI(3)P is produced by the INPP4A hydrolysis of PI(3,4)P2, and this is necessary for actin-driven endocytosis. Both Cdc42·guanosine triphosphate and SNX9 activate N-WASP-WIP- and Arp2/3-mediated actin nucleation. Membrane curvature, PI(4,5)P2, and PI(3)P signals are needed for SNX9 assembly via its PX-BAR domain, whereas signaling through Cdc42 is activated by PI(4,5)P2 alone. INPP4A activity is stimulated by high membrane curvature and synergizes with SNX9 BAR domain binding in a process we call curvature cascade amplification. We show that the SNX9-driven actin comets that arise on human disease-associated oculocerebrorenal syndrome of Lowe (OCRL) deficiencies are reduced by inhibiting PI(3)P production, suggesting PI(3)P kinase inhibitors as a therapeutic strategy in Lowe syndrome.
UR - http://www.scopus.com/inward/record.url?scp=85032893190&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85032893190&partnerID=8YFLogxK
U2 - 10.1083/jcb.201704061
DO - 10.1083/jcb.201704061
M3 - Article
C2 - 28923975
AN - SCOPUS:85032893190
VL - 216
SP - 3745
EP - 3765
JO - Journal of Cell Biology
JF - Journal of Cell Biology
SN - 0021-9525
IS - 11
ER -