A mutant cell line resistant to Vibrio parahaemolyticus thermostable direct hemolysin (TDH): its potential in identification of putative receptor for TDH.
Thermostable direct hemolysin (TDH), a pore-forming toxin produced by Vibrio parahaemolyticus, is cytotoxic to Rat-1, a fibroblast cell line derived from rat embryo. Through mutagenesis of Rat-1 with nitrosoguanidine, we established a mutant cell line, MR-T1. MR-T1 was over 200 times more resistant to the cytotoxic activity of TDH than Rat-1. TDH increased membrane permeability of Rat-1 but not of MR-T1. Binding analysis showed that, while being able to bind to Rat-1. TDH failed to bind to MR-T1, indicating that MR-T1 is deficient in the putative receptor for TDH. Somatic hybrid cells between Rat-1 and MR-T1 were similarly sensitive to TDH as Rat-1. Moreover, TDH could bind to the hybrid cells as well as to Rat-1 cells. These results indicate that MR-T1 is promising for complementation cloning of a gene related to the putative receptor for TDH.
Tang
G
G
Department of Bacterial Infections, Research Institute for Microbial Diseases, Osaka University, Suita, Japan.
Iida
T
T
Inoue
H
H
Yutsudo
M
M
Yamamoto
K
K
Honda
T
T
D000818:Animals; D001427:Bacterial Toxins; D002460:Cell Line / Q000276:immunology; D004351:Drug Resistance / Q000235:genetics; D006460:Hemolysin Proteins / Q000494:pharmacology*; D006822:Hybrid Cells / Q000276:immunology; D016296:Mutagenesis; D051381:Rats; D011956:Receptors, Cell Surface / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D014736:Vibrio parahaemolyticus / Q000276:immunology*
D001427:Bacterial Toxins; D006460:Hemolysin Proteins; D011956:Receptors, Cell Surface; C068489:thermostable direct hemolysin