The ER Tether VAPA is Required for Proper Cell Motility and Anchors ER-PM Contact Sites to Focal Adhesions

Overview

Journal Elife

Specialty Biology

Date 2024 Mar 6

PMID 38446032

Authors

Hugo Siegfried

Georges Farkouh

Remi Le Borgne

Catherine Pioche-Durieu

Thais De Azevedo Laplace

Agathe Verraes

Lucien Daunas

Jean-Marc Verbavatz

Melina L Heuze

Affiliations

Soon will be listed here.

Abstract

Cell motility processes highly depend on the membrane distribution of Phosphoinositides, giving rise to cytoskeleton reshaping and membrane trafficking events. Membrane contact sites serve as platforms for direct lipid exchange and calcium fluxes between two organelles. Here, we show that VAPA, an ER transmembrane contact site tether, plays a crucial role during cell motility. CaCo2 adenocarcinoma epithelial cells depleted for VAPA exhibit several collective and individual motility defects, disorganized actin cytoskeleton and altered protrusive activity. During migration, VAPA is required for the maintenance of PI(4)P and PI(4,5)P2 levels at the plasma membrane, but not for PI(4)P homeostasis in the Golgi and endosomal compartments. Importantly, we show that VAPA regulates the dynamics of focal adhesions (FA) through its MSP domain, is essential to stabilize and anchor ventral ER-PM contact sites to FA, and mediates microtubule-dependent FA disassembly. To conclude, our results reveal unknown functions for VAPA-mediated membrane contact sites during cell motility and provide a dynamic picture of ER-PM contact sites connection with FA mediated by VAPA.

Citing Articles

Molecular characterization underlying IFN-α2 treatment in polycythemia vera: a transcriptomic overview.

Liu F, Li K Mol Cell Biochem. 2025; .

PMID: 40029555 DOI: 10.1007/s11010-025-05238-7.

Allosteric regulation of the tyrosine phosphatase PTP1B by a protein-protein interaction.

Chartier C, Woods V, Xu Y, van Vlimmeren A, Johns A, Jovanovic M Protein Sci. 2024; 34(1):e70016.

PMID: 39723820 PMC: 11670308. DOI: 10.1002/pro.70016.

Imaging and proteomics toolkits for studying organelle contact sites.

Gamuyao R, Chang C Front Cell Dev Biol. 2024; 12:1466915.

PMID: 39381373 PMC: 11458464. DOI: 10.3389/fcell.2024.1466915.

References

Xu W, Wang P, Petri B, Zhang Y, Tang W, Sun L . Integrin-induced PIP5K1C kinase polarization regulates neutrophil polarization, directionality, and in vivo infiltration. Immunity. 2010; 33(3):340-50. PMC: 2947797. DOI: 10.1016/j.immuni.2010.08.015. View

Ezratty E, Partridge M, Gundersen G . Microtubule-induced focal adhesion disassembly is mediated by dynamin and focal adhesion kinase. Nat Cell Biol. 2005; 7(6):581-90. DOI: 10.1038/ncb1262. View

Vance J . Phospholipid synthesis in a membrane fraction associated with mitochondria. J Biol Chem. 1990; 265(13):7248-56. View

Guadagno N, Margiotta A, Bjornestad S, Haugen L, Kjos I, Xu X . Rab18 regulates focal adhesion dynamics by interacting with kinectin-1 at the endoplasmic reticulum. J Cell Biol. 2020; 219(7). PMC: 7337506. DOI: 10.1083/jcb.201809020. View

Chang C, Hsieh T, Yang T, Rothberg K, Azizoglu D, Volk E . Feedback regulation of receptor-induced Ca2+ signaling mediated by E-Syt1 and Nir2 at endoplasmic reticulum-plasma membrane junctions. Cell Rep. 2013; 5(3):813-25. DOI: 10.1016/j.celrep.2013.09.038. View

Hernandez M, Sala M, Balsamo J, Lilien J, Arregui C . ER-bound PTP1B is targeted to newly forming cell-matrix adhesions. J Cell Sci. 2006; 119(Pt 7):1233-43. DOI: 10.1242/jcs.02846. View

Lacalle R, Peregil R, Albar J, Merino E, Martinez-A C, Merida I . Type I phosphatidylinositol 4-phosphate 5-kinase controls neutrophil polarity and directional movement. J Cell Biol. 2007; 179(7):1539-53. PMC: 2373511. DOI: 10.1083/jcb.200705044. View

P F Nader G, Ezratty E, Gundersen G . FAK, talin and PIPKIγ regulate endocytosed integrin activation to polarize focal adhesion assembly. Nat Cell Biol. 2016; 18(5):491-503. DOI: 10.1038/ncb3333. View

Daste F, Walrant A, Holst M, Gadsby J, Mason J, Lee J . Control of actin polymerization via the coincidence of phosphoinositides and high membrane curvature. J Cell Biol. 2017; 216(11):3745-3765. PMC: 5674896. DOI: 10.1083/jcb.201704061. View

10.

Stefan C, Manford A, Baird D, Yamada-Hanff J, Mao Y, Emr S . Osh proteins regulate phosphoinositide metabolism at ER-plasma membrane contact sites. Cell. 2011; 144(3):389-401. DOI: 10.1016/j.cell.2010.12.034. View

11.

Hammond G, Machner M, Balla T . A novel probe for phosphatidylinositol 4-phosphate reveals multiple pools beyond the Golgi. J Cell Biol. 2014; 205(1):113-26. PMC: 3987136. DOI: 10.1083/jcb.201312072. View

12.

Mao D, Lin G, Tepe B, Zuo Z, Tan K, Senturk M . VAMP associated proteins are required for autophagic and lysosomal degradation by promoting a PtdIns4P-mediated endosomal pathway. Autophagy. 2019; 15(7):1214-1233. PMC: 6613884. DOI: 10.1080/15548627.2019.1580103. View

13.

Idevall-Hagren O, Dickson E, Hille B, Toomre D, De Camilli P . Optogenetic control of phosphoinositide metabolism. Proc Natl Acad Sci U S A. 2012; 109(35):E2316-23. PMC: 3435206. DOI: 10.1073/pnas.1211305109. View

14.

Valon L, Marin-Llaurado A, Wyatt T, Charras G, Trepat X . Optogenetic control of cellular forces and mechanotransduction. Nat Commun. 2017; 8:14396. PMC: 5309899. DOI: 10.1038/ncomms14396. View

15.

Kim S, Kedan A, Marom M, Gavert N, Keinan O, Selitrennik M . The phosphatidylinositol-transfer protein Nir2 binds phosphatidic acid and positively regulates phosphoinositide signalling. EMBO Rep. 2013; 14(10):891-9. PMC: 3807235. DOI: 10.1038/embor.2013.113. View

16.

Altan-Bonnet N, Sougrat R, Liu W, Snapp E, Ward T, Lippincott-Schwartz J . Golgi inheritance in mammalian cells is mediated through endoplasmic reticulum export activities. Mol Biol Cell. 2005; 17(2):990-1005. PMC: 1356606. DOI: 10.1091/mbc.e05-02-0155. View

17.

Dong J, Du X, Wang H, Wang J, Lu C, Chen X . Allosteric enhancement of ORP1-mediated cholesterol transport by PI(4,5)P/PI(3,4)P. Nat Commun. 2019; 10(1):829. PMC: 6381110. DOI: 10.1038/s41467-019-08791-0. View

18.

Alpy F, Rousseau A, Schwab Y, Legueux F, Stoll I, Wendling C . STARD3 or STARD3NL and VAP form a novel molecular tether between late endosomes and the ER. J Cell Sci. 2013; 126(Pt 23):5500-12. DOI: 10.1242/jcs.139295. View

19.

Murphy S, Levine T . VAP, a Versatile Access Point for the Endoplasmic Reticulum: Review and analysis of FFAT-like motifs in the VAPome. Biochim Biophys Acta. 2016; 1861(8 Pt B):952-961. DOI: 10.1016/j.bbalip.2016.02.009. View

20.

Lev S, Ben Halevy D, Peretti D, Dahan N . The VAP protein family: from cellular functions to motor neuron disease. Trends Cell Biol. 2008; 18(6):282-90. DOI: 10.1016/j.tcb.2008.03.006. View