Spatial proximity of proteins surrounding zyxin under force-bearing conditions
| Autor: | Volkmar Heinrich, Kyle A. Jacobs, Joleen S Cheah, Jacqueline L Yee, Shuji Ueda, Tzu Wei Lai, Reca Caballelo, Soichiro Yamada |
|---|---|
| Přispěvatelé: | Kaibuchi, Kozo |
| Rok vydání: | 2021 |
| Předmět: |
1.1 Normal biological development and functioning
macromolecular substances Biology Stress Medical and Health Sciences Madin Darby Canine Kidney Cells Zyxin law.invention 03 medical and health sciences chemistry.chemical_compound Dogs 0302 clinical medicine Biotin Underpinning research law Animals Molecular Biology Mechanical Phenomena 030304 developmental biology chemistry.chemical_classification Focal Adhesions 0303 health sciences DNA ligase Bearing (mechanical) Brief Report Microfilament Proteins Cell Biology Biological Sciences Mechanical Phosphoproteins Actins Biomechanical Phenomena Cell biology Actin Cytoskeleton chemistry Biotinylation Stress Mechanical Cell Adhesion Molecules 030217 neurology & neurosurgery Developmental Biology |
| Zdroj: | Molecular Biology of the Cell Molecular biology of the cell, vol 32, iss 13 |
| ISSN: | 1939-4586 1059-1524 |
| DOI: | 10.1091/mbc.e19-10-0568 |
| Popis: | Sensing physical forces is a critical first step in mechano-transduction of cells. Zyxin, a LIM domain-containing protein, is recruited to force-bearing actin filaments and is thought to repair and strengthen them. Yet, the precise force-induced protein interactions surrounding zyxin remain unclear. Using BioID analysis, we identified proximal proteins surrounding zyxin under normal and force-bearing conditions by label-free mass spectrometry analysis. Under force-bearing conditions, increased biotinylation of α-actinin 1, α-actinin 4, and AFAP1 were detected, and these proteins accumulated along force-bearing actin fibers independently from zyxin, albeit at a lower intensity than zyxin. VASP also accumulated along force-bearing actin fibers in a zyxin-dependent manner, but the biotinylation of VASP remained constant regardless of force, supporting the model of a free zyxin-VASP complex in the cytoplasm being corecruited to tensed actin fibers. In addition, ARHGAP42, a RhoA GAP, was also identified as a proximal protein of zyxin and colocalized with zyxin along contractile actin bundles. The overexpression of ARHGAP42 reduced the rate of small wound closure, a zyxin-dependent process. These results demonstrate that the application of proximal biotinylation can resolve the proximity and composition of protein complexes as a function of force, which had not been possible with traditional biochemical analysis. |
| Databáze: | OpenAIRE |
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