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Constitutive active GTPases Rac and Cdc42 are associated with endoreplication in PAE cells.
D F R Muris1, T Verschoor, N Divecha
1Division of Tumour Biology, The Netherlands Cancer Institute, Plesmanlaan 121, Amsterdam, The Netherlands.
European Journal of Cancer (Oxford, England : 1990)
|August 15, 2002
Summary
Constitutively active Rac and Cdc42 GTPases induce cell cycle errors, leading to polyploid cells and genomic instability. This phenomenon, termed mitotic slippage, involves DNA replication without cell division.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Rho-like GTPases regulate fundamental cellular processes like motility and cytoskeletal organization.
- Activation of Rho-like GTPases by extracellular signals influences cell behavior.
Purpose of the Study:
- To investigate the impact of constitutively active Rac and Cdc42 variants on cell cycle progression.
- To elucidate the mechanisms underlying cell cycle transition disruptions caused by these GTPases.
Main Methods:
- Utilized isopropylthiogalactoside (IPTG)-inducible transfectants of porcine aortic endothelial (PAE) cells expressing WtRac, Cdc42, V12Rac, and V12Cdc42.
- Employed fluorescent activated cell sorter (FACS) analysis and bromodeoxyuridine (BrdU) incorporation assays.
Main Results:
- Expression of V12Rac or V12Cdc42 led to G2/M phase enrichment and the formation of multinucleated, polyploid cells.
- These cells exhibited genomic instability and eventual detachment from the culture plate.
- Evidence of endoreplication occurring independently of cytokinesis was observed.
Conclusions:
- Induction of constitutively active V12Rac and V12Cdc42 triggers 'mitotic slippage'.
- Mitotic slippage involves endoreplication and genomic instability, disrupting normal cell cycle progression.
- Dysregulation of Rho-like GTPases can lead to significant cellular abnormalities.