Inhibition of small G proteins by clostridium sordellii lethal toxin activates cdc2 and MAP kinase in Xenopus oocytes

• Published in: Developmental biology 1998-12, Vol.204 (2), p.592-602
• Main Authors: Rime, H, Talbi, N, Popoff, M R, Suziedelis, K, Jessus, C, Ozon, R
• Format: Article
• Language: English
• Subjects: Animals; Bacterial Toxins - toxicity; Calcium-Calmodulin-Dependent Protein Kinases - metabolism; CDC2 Protein Kinase - metabolism; Clostridium; Development Biology; Enzyme Activation - drug effects; Female; GTP-Binding Proteins - antagonists & inhibitors; Life Sciences; Oocytes - drug effects; Oocytes - metabolism; Signal Transduction - drug effects; Xenopus
• ISSN: 0012-1606; 1095-564X
• DOI: 10.1006/dbio.1998.9069

The lethal toxin (LT) from Clostridium sordellii is a glucosyltransferase that modifies and inhibits small G proteins of the Ras family, Ras and Rap, as well as Rac proteins. LT induces cdc2 kinase activation and germinal vesicle breakdown (GVBD) when microinjected into full-grown Xenopus oocytes. Toxin B from Clostridium difficile, that glucosylates and inactivates Rac proteins, does not induce cdc2 activation, indicating that proteins of the Ras family, Ras and/or Rap, negatively regulate cdc2 kinase activation in Xenopus oocyte. In oocyte extracts, LT catalyzes the incorporation of [14C]glucose into a group of proteins of 23 kDa and into one protein of 27 kDa. The 23-kDa proteins are recognized by anti-Rap1 and anti-Rap2 antibodies, whereas the 27-kDa protein is recognized by several anti-Ras antibodies and probably corresponds to K-Ras. Microinjection of LT into oocytes together with UDP-[14C]glucose results in a glucosylation pattern similar to the in vitro glucosylation, indicating that the 23- and 27-kDa proteins are in vivo substrates of LT. In vivo time-course analysis reveals that the 27-kDa protein glucosylation is completed within 2 h, well before cdc2 kinase activation, whereas the 23-kDa proteins are partially glucosylated at GVBD. This observation suggests that the 27-kDa Ras protein could be the in vivo target of LT allowing cdc2 kinase activation. Interestingly, inactivation of Ras proteins does not prevent the phosphorylation of c-Raf1 and the activation of MAP kinase that occurs normally around GVBD.