Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307, Dresden, Germany.
Division of Cell Biology, Life Science Institute, Kurume University, Hyakunen-Kohen 1-1, Kurume, Fukuoka, 839-0864, Japan.
Nat Commun. 2017 Nov 2;8(1):1266. doi: 10.1038/s41467-017-01147-6.
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.
为了分裂，大多数动物细胞会剧烈改变形状并在细胞外限制下聚集。有丝分裂细胞通过产生细胞内压力来促进这个过程，收缩的肌动球蛋白皮层将其引导为形状。在这里，我们引入了一种基于基因组规模的微悬臂梁和 RNAi 的方法，以表型分析超过 1000 个基因对有丝分裂细胞在限制下聚集的贡献。我们的筛选分析了有丝分裂细胞的聚集力、压力和体积，并定位了选定的蛋白质。我们确定了 49 个与有丝分裂聚集相关的基因，其中很大一部分以前与有丝分裂或细胞力学无关。在这些基因中，耗尽内质网定位蛋白 FAM134A 会通过使皮质肌球蛋白 II 去定位来影响中期板的对齐和压力产生，从而损害有丝分裂的进行。此外，沉默 DJ-1 基因揭示了与线粒体相关的帕金森病和有丝分裂压力之间的联系。我们得出结论，机械表型分析是研究控制细胞形状的机制的有力方法。
