Over-expression of GAPDH induces apoptosis in COS-7 cells transfected with cloned GAPDH cDNAs.
The cDNAs for glyceraldehyde-3-phosphate dehydrogenase (GAPDH; EC 1.2.1.12) were cloned from cerebellar neurons undergoing age-induced apoptosis and/or healthy cells. COS-7 cells were transfected with the isolated GAPDH cDNAs using to the Lipofectamine method. Assessment of cell death in this paradigm was performed by monitoring the co-transfected luciferase activities and the characterization of cell death was examined by the DNA fragmentation assay and Hoechst dye nuclear staining. These observations show that over-expression of GAPDH occurring from both cDNAs robustly induces apoptotic death in the transfected COS-7 cell cultures. Confocal-immunocytochemical studies using this GAPDH-specific monoclonal antibody revealed that nuclear translocation of overexpressed GAPDH is a primary apoptotic event. Our results directly demonstrate that over-expressed GAPDH functions as a 'killing protein' in apoptosis.
Tajima
H
H
Minase Research Institute, Ono Pharmaceutical Company Ltd., Osaka, Japan.
Tsuchiya
K
K
Yamada
M
M
Kondo
K
K
Katsube
N
N
Ishitani
R
R
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D019556:COS Cells; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D005786:Gene Expression Regulation / Q000502:physiology*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection
D018076:DNA, Complementary; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases