title: Multiparametric Analysis of Cell Shape Demonstrates that β-PIX Directly Couples YAP Activation to Extracellular Matrix Adhesion.
abstract: Mechanical signals from the extracellular matrix (ECM) and cellular geometry regulate the nuclear translocation of transcriptional regulators such as Yes-associated protein (YAP). Elucidating how physical signals control the activity of mechanosensitive proteins poses a technical challenge, because perturbations that affect cell shape may also affect protein localization indirectly. Here, we present an approach that mitigates confounding effects of cell-shape changes, allowing us to identify direct regulators of YAP localization. This method uses single-cell image analysis and statistical models that exploit the naturally occurring heterogeneity of cellular populations. Through systematic depletion of all human kinases, Rho family GTPases, GEFs, and GTPase activating proteins (GAPs), together with targeted chemical perturbations, we found that β-PIX, a Rac1/Ccd42 GEF, and PAK2, a Rac1/Cdc42 effector, drive both YAP activation and cell-ECM adhesion turnover during cell spreading. Our observations suggest that coupling YAP to adhesion dynamics acts as a mechano-timer, allowing cells to rapidly tune gene expression in response to physical signals.
authors.affiliation: Chester Beatty Laboratories, Division of Cancer Biology, Institute of Cancer Research, 237 Fulham Road, London SW3 6JB, UK. Electronic address: juliasero@post.harvard.edu.
authors.forename: Julia E
authors.initials: JE
authors.lastname: Sero
authors.affiliation: Chester Beatty Laboratories, Division of Cancer Biology, Institute of Cancer Research, 237 Fulham Road, London SW3 6JB, UK.
authors.forename: Chris
authors.initials: C
authors.lastname: Bakal
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology; D018797:Cell Cycle Proteins / Q000235:genetics; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology; D048430:Cell Shape / Q000502:physiology*; D005109:Extracellular Matrix / Q000378:metabolism / Q000502:physiology; D005260:Female; D020690:GTPase-Activating Proteins / Q000235:genetics; D006801:Humans; D009687:Nuclear Proteins / Q000502:physiology*; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D064067:Rho Guanine Nucleotide Exchange Factors / Q000502:physiology*; D015398:Signal Transduction; D059010:Single-Cell Analysis / Q000379:methods; D014157:Transcription Factors / Q000502:physiology*; D020764:cdc42 GTP-Binding Protein / Q000235:genetics / Q000502:physiology; D020830:rac1 GTP-Binding Protein / Q000235:genetics / Q000502:physiology; D020742:rhoA GTP-Binding Protein / Q000235:genetics
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D018797:Cell Cycle Proteins; D020690:GTPase-Activating Proteins; D009687:Nuclear Proteins; D064067:Rho Guanine Nucleotide Exchange Factors; D014157:Transcription Factors; C456659:YY1AP1 protein, human; D020764:cdc42 GTP-Binding Protein; D020830:rac1 GTP-Binding Protein; D020742:rhoA GTP-Binding Protein
keywords: Cdc42; FAK; Rac1; YAP; adhesion; beta-PIX; breast cancer; cell shape; mechanobiology; morphology