title: Cell cortex composition and homeostasis resolved by integrating proteomics and quantitative imaging.
abstract: The cellular actin cortex is the cytoskeletal structure primarily responsible for the control of animal cell shape and as such plays a central role in cell division, migration, and tissue morphogenesis. Due to the lack of experimental systems where the cortex can be investigated independently from other organelles, little is known about its composition, assembly, and homeostasis. Here, we describe novel tools to resolve the composition and regulation of the cortex. We report and validate a protocol for cortex purification based on the separation of cellular blebs. Mass spectrometry analysis of purified cortices provides a first extensive list of cortical components. To assess the function of identified proteins, we design an automated imaging assay for precise quantification of cortical actomyosin assembly dynamics. We show subtle changes in cortex assembly dynamics upon depletion of the identified cortical component profilin. Our widely applicable integrated method paves the way for systems-level investigations of the actomyosin cortex and its regulation during morphogenesis.
authors.affiliation: Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany; International Institute of Molecular and Cell Biology, Warsaw, Poland.
authors.forename: Maté
authors.initials: M
authors.lastname: Biro
authors.forename: Yves
authors.initials: Y
authors.lastname: Romeo
authors.forename: Sonja
authors.initials: S
authors.lastname: Kroschwald
authors.forename: Miia
authors.initials: M
authors.lastname: Bovellan
authors.forename: Annett
authors.initials: A
authors.lastname: Boden
authors.forename: Joseph
authors.initials: J
authors.lastname: Tcherkezian
authors.forename: Philippe P
authors.initials: PP
authors.lastname: Roux
authors.forename: Guillaume
authors.initials: G
authors.lastname: Charras
authors.forename: Ewa K
authors.initials: EK
authors.lastname: Paluch
mesh_terms: D000199:Actins / Q000378:metabolism*; D000205:Actomyosin / Q000378:metabolism; D022082:Cellular Structures / Q000378:metabolism* / Q000648:ultrastructure; D006367:HeLa Cells; D006706:Homeostasis; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D051304:Profilins / Q000378:metabolism; D040901:Proteomics / Q000379:methods*
chemical_list: D000199:Actins; D051304:Profilins; D000205:Actomyosin
keywords: actin assembly; actin cortex; blebs; proteomics; quantitative imaging