title: Cellular stability of Rho-GTPases glucosylated by Clostridium difficile toxin B.
abstract: Mono-glucosylation of Rho, Rac, and Cdc42 by Clostridium difficile toxin B (TcdB) induces changes of actin dynamics and apoptosis. When fibroblasts were treated with TcdB, an apparent decrease of the cellular Rac1 level was observed when applying anti-Rac1(Mab 102). This decrease was not based on degradation as inhibition of the proteasome by lactacystin did not stabilise cellular Rac1 levels. The application of anti-Rac1 (Mab 23A8) showed that the cellular Rac1 level slightly increased in TcdB-treated fibroblasts; thus, the apparent loss of cellular Rac1 was not due to degradation but due to impaired recognition of glucosylated Rac1 by anti-Rac1 (Mab 102). In contrast, recognition of RhoA by anti-RhoA (Mab 26C4) and Cdc42 by anti-Cdc42 (Mab 44) was not altered by glucosylation; a transient decrease of cellular RhoA and Cdc42 in TcdB-treated fibroblasts was indeed due to proteasomal degradation, as inhibition of the proteasome by lactacystin stabilised both cellular RhoA and Cdc42 levels. The finding that the apparent decrease of Rac1 reflects Rac1 glucosylation offers a valuable tool to determine Rac1 glucosylation.
authors.affiliation: Department of Toxicology, Hannover Medical School, Germany. genth.harald@mh-hannover.de
authors.forename: Harald
authors.initials: H
authors.lastname: Genth
authors.forename: Johannes
authors.initials: J
authors.lastname: Huelsenbeck
authors.forename: Birgit
authors.initials: B
authors.lastname: Hartmann
authors.forename: Fred
authors.initials: F
authors.lastname: Hofmann
authors.forename: Ingo
authors.initials: I
authors.lastname: Just
authors.forename: Ralf
authors.initials: R
authors.lastname: Gerhard
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism* / Q000494:pharmacology; D001427:Bacterial Toxins / Q000378:metabolism* / Q000494:pharmacology; D002474:Cell-Free System; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006031:Glycosylation; D051379:Mice; D041681:NIH 3T3 Cells; D020741:rho GTP-Binding Proteins / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; C031815:toxB protein, Clostridium difficile; D020741:rho GTP-Binding Proteins