title: A mutant cell line resistant to Vibrio parahaemolyticus thermostable direct hemolysin (TDH): its potential in identification of putative receptor for TDH.
abstract: 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.
authors.affiliation: Department of Bacterial Infections, Research Institute for Microbial Diseases, Osaka University, Suita, Japan.
authors.forename: G
authors.initials: G
authors.lastname: Tang
authors.forename: T
authors.initials: T
authors.lastname: Iida
authors.forename: H
authors.initials: H
authors.lastname: Inoue
authors.forename: M
authors.initials: M
authors.lastname: Yutsudo
authors.forename: K
authors.initials: K
authors.lastname: Yamamoto
authors.forename: T
authors.initials: T
authors.lastname: Honda
mesh_terms: 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*
chemical_list: D001427:Bacterial Toxins; D006460:Hemolysin Proteins; D011956:Receptors, Cell Surface; C068489:thermostable direct hemolysin