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Cell expression of a four extra octarepeat mutated PrPC modifies cell structure and cell cycle regulation
Sergio F Martín1, María E Herva, Juan-Carlos Espinosa
1Centro de Investigación en Sanidad (CISA-INIA), Ctra. de Algete a El Casar, km. 8.100, 28130 Valdeolmos, Madrid, Spain.
FEBS Letters
|July 11, 2006
Summary
The bovine PrP(C) mutation (Bo-10ORPrP(C)) in RK13 cells causes cell cycle arrest at G2/M, impacting cell growth and morphology. Proteasome inhibition amplifies this effect, suggesting PrP(C) involvement in cell cycle regulation.
Area of Science:
- Cell Biology
- Neuroscience
- Biochemistry
Background:
- Prion protein cellular (PrP(C)) function is not fully understood.
- Mutations in PrP(C) are linked to neurodegenerative diseases.
- Investigating the cellular impact of PrP(C) mutations is crucial.
Purpose of the Study:
- To investigate the cellular effects of a bovine PrP(C) octarepeat insertional mutation (Bo-10ORPrP(C)).
- To determine the role of PrP(C) in cell morphology and cell cycle regulation.
- To explore the interaction between PrP(C) and proteasome function.
Main Methods:
- Generation of RK13 cell lines expressing wild-type (Bo-6ORPrP(C)) and mutant (Bo-10ORPrP(C)) bovine PrP(C).
- Assessment of cell growth rates, PrP(C) solubility, cell size, and actin cytoskeleton organization.
- Treatment with Clostridium difficile toxin-B and proteasome inhibitors.
- Cell cycle analysis using flow cytometry.
Main Results:
- Bo-10ORPrP(C) expression led to partially insoluble PrP(C), reduced cell growth, altered cell size, and actin cytoskeleton reorganization.
- Clostridium difficile toxin-B reversed actin cytoskeleton changes.
- Cells expressing Bo-10ORPrP(C) exhibited increased G2/M cell cycle arrest.
- Proteasome inhibitors exacerbated G2/M arrest, particularly in Bo-10ORPrP(C) expressing cells.
Conclusions:
- Bovine PrP(C) with a four extra octarepeat insertional mutation induces cell cycle arrest at G2/M in RK13 cells.
- PrP(C) plays a role in regulating cell morphology and cell cycle progression.
- The findings provide insights into PrP mutation-associated diseases and potential therapeutic targets.
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