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Neurochemical Research

, Volume 33, Issue 7, pp 1393–1400 | Cite as

Dequalinium-Induced Cell Death of Yeast Expressing α-Synuclein-GFP Fusion Protein

  • In-Hwan Lee
  • Hui-Young Kim
  • Myunghye Kim
  • Ji-Sook Hahn
  • Seung R. PaikEmail author
Original Paper

Abstract

Intracellular toxic effects of the dequalinium-induced protofibrils of α-synuclein have been investigated with the yeast system expressing α-synuclein-GFP fusion protein in single copy, which appears in the green halo around the plasma membrane. Intracellular responses of the green fluorescent protein were analyzed as the cells were treated with dequalinium (DQ) and lactacystin. Yeast cells expressing α-synuclein-GFP were susceptible to both compounds in α-synuclein-dependent manner. Upon DQ treatment, the green halo became smeared throughout the cytoplasm while lactacystin induced a few discrete green dots, reflecting intracellular formation of the protofibrils and the protein inclusions, respectively. The DQ-treated yeast cells were intensely stained with the nucleic acid stains of cell-permeable Hoechst 33342 and cell-impermeable propidium imidione, indicating that nucleus has been disrupted in addition to plasma membrane destabilization. Those DQ-treated yeast cells, however, still contained active mitochondria identified with MitoTracker Red. Therefore, the DQ-induced protofibrillar state of α-synuclein-GFP has been suggested to cause the nuclear damage either independently or in combination with the membrane destabilization without affecting mitochondria.

Keywords

Dequalinium α-Synuclein Yeast Neurodegenerative disorder Cell death Lactacystin Nuclear malfunction Mitochondria dysfunction Parkinson’s disease 

Notes

Acknowledgements

This study was supported in part by a basic research program grant (R01-2007-000-20089-0) from Korea Science and Engineering, a grant from Technology Development Program of the Ministry of Agriculture and Forestry, and by a Grant from the Seoul R&BD Program (10538).

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Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • In-Hwan Lee
    • 1
  • Hui-Young Kim
    • 1
  • Myunghye Kim
    • 1
  • Ji-Sook Hahn
    • 1
  • Seung R. Paik
    • 1
    Email author
  1. 1.School of Chemical and Biological Engineering, College of EngineeringSeoul National UniversitySeoulRepublic of Korea

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