Synergistic phase separation of two pathways promotes integrin clustering and nascent adhesion formation

Abstract
Data availability
Article and author information
Metrics

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 ('Cas') and Focal adhesion kinase ('FAK') 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.

Data availability

If the article is accepted, all imaging data will be deposited in the Dryad database before publication.

The following data sets were generated

1. Case L
2. Rosen M
(2022) Synergistic Phase Separation of Two Pathways Promotes Integrin Clustering and Nascent Adhesion Formation
Dryad Digital Repository, doi:10.5061/dryad.9p8cz8wj0.

https://doi.org/10.5061/dryad.9p8cz8wj0

Article and author information

Author details

Lindsay B Case

Department of Biophysics, Howard Hughes Medical Institute, The University of Texas Southwestern Medical Center, Dallas, United States

For correspondence
lcase@mit.edu

Competing interests
No competing interests declared.
Milagros De Pasquale

Department of Biology, Massachusetts Institute of Technology, Cambridge, United States

Competing interests
No competing interests declared.
Lisa Henry

Department of Biophysics, Howard Hughes Medical Institute, The University of Texas Southwestern Medical Center, Dallas, United States

Competing interests
No competing interests declared.
Michael K Rosen

Department of Biophysics, Howard Hughes Medical Institute, The University of Texas Southwestern Medical Center, Dallas, United States

For correspondence
michael.rosen@utsouthwestern.edu

Competing interests
Michael K Rosen, is a co-founder of Faze Medicines.

"This ORCID iD identifies the author of this article:" 0000-0002-0775-7917

Funding

Damon Runyon Cancer Research Foundation (postdoctoral fellowship,DRG-2249-16)

Lindsay B Case
Michael K Rosen

Damon Runyon Cancer Research Foundation (Dale Frey Scientist Award,DFS-38-20)

Lindsay B Case

Howard Hughes Medical Institute (Investigator)

Michael K Rosen

Welch Foundation (I-1544)

Michael K Rosen

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.