Document Detail


Bcl-2 blocks apoptosis caused by pierisin-1, a guanine-specific ADP-ribosylating toxin from the cabbage butterfly.
MedLine Citation:
PMID:  12147221     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Pierisin-1, a 98-kDa protein that induces apoptosis in mammalian cell lines, is capable of being incorporated into cells where it ADP-ribosylates guanine residues in DNA. To investigate the apoptotic pathway induced by this unique protein, the bcl-2 gene was transfected into HeLa cells. Cy2-fluorescent pierisin-1 was incorporated into the resultant cells expressing Bcl-2 protein and ADP-ribosylated dG was detected to almost the same extent as in parent cells. However, bcl-2-transfected HeLa cells did not display apoptotic morphological changes, PARP cleavage, and DNA fragmentation, indicating acquisition of resistance. In parent HeLa cells, activation of caspase-9 and release of cytochrome c were observed after 8h treatment with 0.5ng/ml pierisin-1. Caspase substrate assays revealed further cleavage of Ac-DEVD-pNA, Ac-VDVAD-pNA, and Ac-VEID-pNA, suggesting activation of caspase-2, -3, and -6 in pierisin-1-treated HeLa cells. The caspase-3 inhibitor, Ac-DEVD-CHO, was also found to inhibit apoptosis. In contrast, this caspase activation was not observed in bcl-2-transfected HeLa cells. Our results thus indicate that pierisin-1-induced apoptosis is mediated primarily via a mitochondrial pathway involving Bcl-2 and caspases.
Authors:
Takashi Kanazawa; Takuo Kono; Masahiko Watanabe; Yuko Matsushima-Hibiya; Tsuyoshi Nakano; Kotaro Koyama; Noriaki Tanaka; Takashi Sugimura; Keiji Wakabayashi
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biochemical and biophysical research communications     Volume:  296     ISSN:  0006-291X     ISO Abbreviation:  Biochem. Biophys. Res. Commun.     Publication Date:  2002 Aug 
Date Detail:
Created Date:  2002-07-30     Completed Date:  2002-09-06     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  0372516     Medline TA:  Biochem Biophys Res Commun     Country:  United States    
Other Details:
Languages:  eng     Pagination:  20-5     Citation Subset:  IM    
Affiliation:
Cancer Prevention Division, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, 104-0045, Tokyo, Japan. tkanazaw@gan2.ncc.go.jp
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MeSH Terms
Descriptor/Qualifier:
ADP Ribose Transferases
Adenosine Diphosphate / metabolism*
Animals
Apoptosis / drug effects,  genetics*
Butterflies
Caspases / metabolism
Cytochrome c Group / metabolism
Enzyme Activation
Genes, bcl-2*
Hela Cells
Humans
Insect Proteins / pharmacology*
Chemical
Reg. No./Substance:
0/Cytochrome c Group; 0/Insect Proteins; 0/pierisin protein, insect; 58-64-0/Adenosine Diphosphate; EC 2.4.2.-/ADP Ribose Transferases; EC 3.4.22.-/Caspases

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