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Characterization of RACK1-depleted mammalian cells by a palette of microscopy approaches reveals defects in cell

Vera Chvalova1, Vignesh Venkadasubramanian1, Zuzana Klimova2

  • 1Laboratory of Cell Signalling, Institute of Microbiology of the Czech Academy of Sciences, Videnska 1083, 142 00, Prague, Czech Republic; Faculty of Science, Charles University, 128 00, Prague, Czech Republic.

Experimental Cell Research
|July 2, 2023
PubMed

Insights

Receptor for Activated C Kinase 1 (RACK1) depletion impairs cell proliferation and causes cell cycle defects in mammalian cells. This study highlights RACK1

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Receptor for Activated C Kinase 1 (RACK1) is an evolutionarily conserved scaffold protein.
  • RACK1 regulates diverse cellular processes.

Purpose of the Study:

  • To investigate the role of RACK1 in mammalian cell proliferation and cell cycle progression.
  • To examine the effects of RACK1 depletion in epithelial and mesenchymal cell lines.

Main Methods:

  • CRISPR/Cas9 and siRNA were used to deplete RACK1 expression in MDCK epithelial cells and Rat2 fibroblasts.
  • Coherence-controlled holographic microscopy, immunofluorescence, and electron microscopy were employed for cell analysis.

Main Results:

  • RACK1 depletion led to decreased cell proliferation.
  • Cells exhibited increased area and perimeter, with the appearance of large binucleated cells.
  • These findings suggest a defect in cell cycle progression.

Conclusions:

  • RACK1 depletion has pleiotropic effects on both epithelial and mesenchymal cells.
  • RACK1 plays an essential role in mammalian cell function.
Keywords:
ActinCell cycleFocal adhesionsMDCK cellsMicroscopy-based analysisMicrovilliRACK1Rat2 cells

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