Genome-scale single-cell mechanical phenotyping reveals disease-related genes involved in mitotic rounding
Cattin, Cédric Joël
Kurzchalia, Teymuras V.
Hyman, Anthony A.
Muller, Daniel J.
- Journal Article
Rights / licenseCreative Commons Attribution 4.0 International
To divide, most animal cells drastically change shape and round up against extracellular confinement. Mitotic cells facilitate this process by generating intracellular pressure, which the contractile actomyosin cortex directs into shape. Here, we introduce a genome-scale microcantilever- and RNAi-based approach to phenotype the contribution of > 1000 genes to the rounding of single mitotic cells against confinement. Our screen analyzes the rounding force, pressure and volume of mitotic cells and localizes selected proteins. We identify 49 genes relevant for mitotic rounding, a large portion of which have not previously been linked to mitosis or cell mechanics. Among these, depleting the endoplasmic reticulum-localized protein FAM134A impairs mitotic progression by affecting metaphase plate alignment and pressure generation by delocalizing cortical myosin II. Furthermore, silencing the DJ-1 gene uncovers a link between mitochondria-associated Parkinson's disease and mitotic pressure. We conclude that mechanical phenotyping is a powerful approach to study the mechanisms governing cell shape Show more
Journal / seriesNature Communications
Organisational unit03870 - Müller, Daniel J.
NotesWe thank M. Düggelin for producing FIB-sculpted cantilevers, J. Helenius and S.P. Ramanathan for help with myosin and actin localization ratio determination, E. Paluch and W. Wurst for cell lines, M. Kuhn for advice on bioinformatics analysis, A. Ssykor and M. Leuschner for transgenic cell production and the MPI-CBG antibody facility for production of the FAM134A antibody, and J. Guck and M. Schlierf for providing access to one of the several AFMs used. This study was supported by the Max Planck Society, by the Swiss National Science Foundation (Advanced Mobility Fellowship to M.P.S. and Grant 310030B_160255 to D.J.M.), the Eidgenössische Technische Hochschule (Research Grant ETH-05 11-2 to D.J.M.), and the NCCR Molecular Systems Engineering.
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