Characterization of RACK1-depleted mammalian cells by a palette of microscopy approaches reveals defects in cell cycle progression and polarity establishment

0574999 - MBÚ 2024 RIV NL eng J - Journal Article
Chvalová, Věra - Venkadasubramanian, Vignesh - Klímová, Zuzana - Vojtová, Jana - Benada, Oldřich - Vaňátko, Ondřej - Vomastek, Tomáš - Groušl, Tomáš
Characterization of RACK1-depleted mammalian cells by a palette of microscopy approaches reveals defects in cell cycle progression and polarity establishment.
Experimental Cell Research. Roč. 430, č. 1 (2023), č. článku 113695. ISSN 0014-4827. E-ISSN 1090-2422
R&D Projects: GA ČR(CZ) GJ19-07603Y; GA ČR GA18-11908S; GA MŠMT(CZ) LUASK22100
Research Infrastructure: Czech-BioImaging II - 90129
Institutional support: RVO:61388971 ; RVO:68378041
Keywords : rack1 * MDCK cells * Rat2 cells * Cell cycle * Microscopy-based analysis * Actin * Focal adhesions * Microvilli
OECD category: Cell biology; Cell biology (UEM-P)
Impact factor: 3.7, year: 2022
Method of publishing: Limited access
https://www.sciencedirect.com/science/article/pii/S0014482723002434?via%3Dihub

The Receptor for Activated C Kinase 1 (RACK1) is an evolutionarily conserved scaffold protein involved in the regulation of numerous cellular processes. Here, we used CRISPR/Cas9 and siRNA to reduce the expression of RACK1 in Madin-Darby Canine Kidney (MDCK) epithelial cells and Rat2 fibroblasts, respectively. RACK1-depleted cells were examined using coherence-controlled holographic microscopy, immunofluorescence, and electron microscopy. RACK1 depletion resulted in decreased cell proliferation, increased cell area and perimeter, and in the appearance of large binucleated cells suggesting a defect in the cell cycle progression. Our results show that the depletion of RACK1 has a pleiotropic effect on both epithelial and mesenchymal cell lines and support its essential role in mammalian cells.
Permanent Link: https://hdl.handle.net/11104/0344913