@article{e142e78845ad4bfc98260cba7f474480,
title = "Synergistic phase separation of two pathways promotes integrin clustering and nascent adhesion formation",
abstract = "Integrin adhesion complexes (IACs) are integrin-based plasma-membrane-associated compartments where cells sense environmental cues. The physical mechanisms and molecular interactions that mediate initial IAC formation are unclear. We found that both p130Cas ({\textquoteleft}Cas{\textquoteright}) and Focal adhesion kinase ({\textquoteleft}FAK{\textquoteright}) undergo liquid-liquid phase separation in vitro under physiologic conditions. Cas-and FAK-driven phase separation is sufficient to reconstitute kindlin-dependent integrin clustering in vitro with recombinant mammalian proteins. In vitro condensates and IACs in mouse embryonic fibroblasts (MEFs) exhibit similar sensitivities to environmental perturbations including changes in temperature and pH. Furthermore, mutations that inhibit or enhance phase separation in vitro reduce or increase the number of IACs in MEFs, respectively. Finally, we find that the Cas and FAK pathways act synergistically to promote phase separation, integrin clustering, IAC formation and partitioning of key components in vitro and in cells. We propose that Cas-and FAK-driven phase separation provides an intracellular trigger for integrin clustering and nascent IAC formation.",
author = "Case, {Lindsay B.} and {De Pasquale}, Milagros and Lisa Henry and Rosen, {Michael K.}",
note = "Funding Information: We thank Steve Hanks (Vanderbilt) and Larisa Ryzhova (Maine Medical Center) for sharing Bcar1-/-cell lines, Robert Liddington (Sanford Burnham Institute) for sharing talin and FAK DNA plasmids, W Todd Miller (Stony Brook University) for sharing p130Cas DNA plasmid, Jun Qin (Cleveland Clinic) for sharing kindlin DNA plasmid, and David Corey and Jiaxin Hu (UT Southwestern Medical Center) for use of their Cary 100 UV-Visible spectrophotometer and Peltier thermal controller. We thank the UT Southwestern Proteomics Core Facility and the Koch Institute Biopolymers & Proteomics Facility for performing mass spectrometry analysis. L.B.C. was a Robert Black Fellow of the Damon Runyon Cancer Research Foundation (DRG-2249?16) and is further supported (in part) by the Damon Runyon Cancer Research Foundation (DFS-38?20). This work was also supported by the Howard Hughes Medical Institute and the Welch Foundation (grant I-1544). Funding Information: We thank Steve Hanks (Vanderbilt) and Larisa Ryzhova (Maine Medical Center) for sharing Bcar1-/-cell lines, Robert Liddington (Sanford Burnham Institute) for sharing talin and FAK DNA plasmids, W Todd Miller (Stony Brook University) for sharing p130Cas DNA plasmid, Jun Qin (Cleveland Clinic) for sharing kindlin DNA plasmid, and David Corey and Jiaxin Hu (UT Southwestern Medical Center) for use of their Cary 100 UV-Visible spectrophotometer and Peltier thermal controller. We thank the UT Southwestern Proteomics Core Facility and the Koch Institute Biopolymers & Proteomics Facility for performing mass spectrometry analysis. L.B.C. was a Robert Black Fellow of the Damon Runyon Cancer Research Foundation (DRG-2249–16) and is further supported (in part) by the Damon Runyon Cancer Research Foundation (DFS-38–20). This work was also supported by the Howard Hughes Medical Institute and the Welch Foundation (grant I-1544). Publisher Copyright: {\textcopyright} Case et al.",
year = "2022",
month = jan,
doi = "10.7554/eLife.72588",
language = "English (US)",
volume = "11",
journal = "eLife",
issn = "2050-084X",
publisher = "eLife Sciences Publications",
}