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J Clin Invest. 1995 March; 95(3): 1026–1031.
PMCID: PMC441436

The low molecular mass GTP-binding protein Rho is affected by toxin A from Clostridium difficile.

Abstract

Enterotoxin A is one of the major virulence factors of Clostridium difficile, and the causative agent of antibiotic-associated pseudomembranous colitis. In cell culture (NIH-3T3, rat basophilic leukemia cells) toxin A inhibits Clostridium botulinum ADP-ribosyltransferase C3 (C3)-catalyzed ADP-ribosylation of the low molecular mass GTP-binding Rho proteins. Rho participates in the regulation of the microfilament cytoskeleton. Decrease in ADP-ribosylation of Rho occurs in a time- and concentration-dependent manner and precedes the toxin A-induced destruction of the actin cytoskeleton. Action of toxin A is not due to proteolytical degradation of Rho or to an inherent ADP-ribosyltransferase activity of toxin A. Toxin A-induced decrease in ADP-ribosylation is observed also in cell lysates and with recombinant RhoA protein. A heat stable low molecular mass cytosolic factor is essential for the toxin effect on Rho. Thus, the enterotoxin (toxin A) resembles the effects of the C. difficile cytotoxin (toxin B) on Rho proteins (Just, I., G. Fritz, K. Aktories, M. Giry, M. R. Popoff, P. Boquet, S. Hegenbath, and C. Von Eichel-Streiber. 1994. J. Biol. Chem. 269:10706-10712). The data indicate that despite different in vivo effects, toxin A and toxin B act on the same cellular target protein Rho to elicit their toxic effects.

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  • Lyerly DM, Krivan HC, Wilkins TD. Clostridium difficile: its disease and toxins. Clin Microbiol Rev. 1988 Jan;1(1):1–18. [PMC free article] [PubMed]
  • Lyerly DM, Lockwood DE, Richardson SH, Wilkins TD. Biological activities of toxins A and B of Clostridium difficile. Infect Immun. 1982 Mar;35(3):1147–1150. [PMC free article] [PubMed]
  • Lyerly DM, Saum KE, MacDonald DK, Wilkins TD. Effects of Clostridium difficile toxins given intragastrically to animals. Infect Immun. 1985 Feb;47(2):349–352. [PMC free article] [PubMed]
  • Triadafilopoulos G, Pothoulakis C, O'Brien MJ, LaMont JT. Differential effects of Clostridium difficile toxins A and B on rabbit ileum. Gastroenterology. 1987 Aug;93(2):273–279. [PubMed]
  • Arnon SS, Mills DC, Day PA, Henrickson RV, Sullivan NM, Wilkins TD. Rapid death of infant rhesus monkeys injected with Clostridium difficile toxins A and B: physiologic and pathologic basis. J Pediatr. 1984 Jan;104(1):34–40. [PubMed]
  • von Eichel-Streiber C, Sauerborn M. Clostridium difficile toxin A carries a C-terminal repetitive structure homologous to the carbohydrate binding region of streptococcal glycosyltransferases. Gene. 1990 Nov 30;96(1):107–113. [PubMed]
  • Barroso LA, Wang SZ, Phelps CJ, Johnson JL, Wilkins TD. Nucleotide sequence of Clostridium difficile toxin B gene. Nucleic Acids Res. 1990 Jul 11;18(13):4004–4004. [PMC free article] [PubMed]
  • Henriques B, Florin I, Thelestam M. Cellular internalisation of Clostridium difficile toxin A. Microb Pathog. 1987 Jun;2(6):455–463. [PubMed]
  • Florin I, Thelestam M. Lysosomal involvement in cellular intoxication with Clostridium difficile toxin B. Microb Pathog. 1986 Aug;1(4):373–385. [PubMed]
  • Nitta AT, Iseman MD, Newell JD, Madsen LA, Goble M. Ten-year experience with artificial pneumoperitoneum for end-stage, drug-resistant pulmonary tuberculosis. Clin Infect Dis. 1993 Feb;16(2):219–222. [PubMed]
  • Fiorentini C, Arancia G, Paradisi S, Donelli G, Giuliano M, Piemonte F, Mastrantonio P. Effects of Clostridium difficile toxins A and B on cytoskeleton organization in HEp-2 cells: a comparative morphological study. Toxicon. 1989;27(11):1209–1218. [PubMed]
  • Ottlinger ME, Lin S. Clostridium difficile toxin B induces reorganization of actin, vinculin, and talin in cultured cells. Exp Cell Res. 1988 Jan;174(1):215–229. [PubMed]
  • Mitchell MJ, Laughon BE, Lin S. Biochemical studies on the effect of Clostridium difficile toxin B on actin in vivo and in vitro. Infect Immun. 1987 Jul;55(7):1610–1615. [PMC free article] [PubMed]
  • Fiorentini C, Thelestam M. Clostridium difficile toxin A and its effects on cells. Toxicon. 1991;29(6):543–567. [PubMed]
  • Kelly CP, Pothoulakis C, LaMont JT. Clostridium difficile colitis. N Engl J Med. 1994 Jan 27;330(4):257–262. [PubMed]
  • Chardin P, Boquet P, Madaule P, Popoff MR, Rubin EJ, Gill DM. The mammalian G protein rhoC is ADP-ribosylated by Clostridium botulinum exoenzyme C3 and affects actin microfilaments in Vero cells. EMBO J. 1989 Apr;8(4):1087–1092. [PubMed]
  • Wiegers W, Just I, Müller H, Hellwig A, Traub P, Aktories K. Alteration of the cytoskeleton of mammalian cells cultured in vitro by Clostridium botulinum C2 toxin and C3 ADP-ribosyltransferase. Eur J Cell Biol. 1991 Apr;54(2):237–245. [PubMed]
  • Aktories K, Rösener S, Blaschke U, Chhatwal GS. Botulinum ADP-ribosyltransferase C3. Purification of the enzyme and characterization of the ADP-ribosylation reaction in platelet membranes. Eur J Biochem. 1988 Mar 1;172(2):445–450. [PubMed]
  • Rubin EJ, Gill DM, Boquet P, Popoff MR. Functional modification of a 21-kilodalton G protein when ADP-ribosylated by exoenzyme C3 of Clostridium botulinum. Mol Cell Biol. 1988 Jan;8(1):418–426. [PMC free article] [PubMed]
  • Nemoto Y, Namba T, Kozaki S, Narumiya S. Clostridium botulinum C3 ADP-ribosyltransferase gene. Cloning, sequencing, and expression of a functional protein in Escherichia coli. J Biol Chem. 1991 Oct 15;266(29):19312–19319. [PubMed]
  • Just I, Mohr C, Schallehn G, Menard L, Didsbury JR, Vandekerckhove J, van Damme J, Aktories K. Purification and characterization of an ADP-ribosyltransferase produced by Clostridium limosum. J Biol Chem. 1992 May 25;267(15):10274–10280. [PubMed]
  • Just I, Schallehn G, Aktories K. ADP-ribosylation of small GTP-binding proteins by Bacillus cereus. Biochem Biophys Res Commun. 1992 Mar 31;183(3):931–936. [PubMed]
  • Sugai M, Hashimoto K, Kikuchi A, Inoue S, Okumura H, Matsumoto K, Goto Y, Ohgai H, Moriishi K, Syuto B, et al. Epidermal cell differentiation inhibitor ADP-ribosylates small GTP-binding proteins and induces hyperplasia of epidermis. J Biol Chem. 1992 Feb 5;267(4):2600–2604. [PubMed]
  • Ridley AJ, Hall A. The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell. 1992 Aug 7;70(3):389–399. [PubMed]
  • Stasia MJ, Jouan A, Bourmeyster N, Boquet P, Vignais PV. ADP-ribosylation of a small size GTP-binding protein in bovine neutrophils by the C3 exoenzyme of Clostridium botulinum and effect on the cell motility. Biochem Biophys Res Commun. 1991 Oct 31;180(2):615–622. [PubMed]
  • Takaishi K, Kikuchi A, Kuroda S, Kotani K, Sasaki T, Takai Y. Involvement of rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI) in cell motility. Mol Cell Biol. 1993 Jan;13(1):72–79. [PMC free article] [PubMed]
  • Hirata K, Kikuchi A, Sasaki T, Kuroda S, Kaibuchi K, Matsuura Y, Seki H, Saida K, Takai Y. Involvement of rho p21 in the GTP-enhanced calcium ion sensitivity of smooth muscle contraction. J Biol Chem. 1992 May 5;267(13):8719–8722. [PubMed]
  • Isomura M, Kaibuchi K, Yamamoto T, Kawamura S, Katayama M, Takai Y. Partial purification and characterization of GDP dissociation stimulator (GDS) for the rho proteins from bovine brain cytosol. Biochem Biophys Res Commun. 1990 Jun 15;169(2):652–659. [PubMed]
  • Yamamoto J, Kikuchi A, Ueda T, Ohga N, Takai Y. A GTPase-activating protein for rhoB p20, a ras p21-like GTP-binding protein--partial purification, characterization and subcellular distribution in rat brain. Brain Res Mol Brain Res. 1990 Jul;8(2):105–111. [PubMed]
  • Morii N, Kawano K, Sekine A, Yamada T, Narumiya S. Purification of GTPase-activating protein specific for the rho gene products. J Biol Chem. 1991 Apr 25;266(12):7646–7650. [PubMed]
  • Fukumoto Y, Kaibuchi K, Hori Y, Fujioka H, Araki S, Ueda T, Kikuchi A, Takai Y. Molecular cloning and characterization of a novel type of regulatory protein (GDI) for the rho proteins, ras p21-like small GTP-binding proteins. Oncogene. 1990 Sep;5(9):1321–1328. [PubMed]
  • Bourmeyster N, Stasia MJ, Garin J, Gagnon J, Boquet P, Vignais PV. Copurification of rho protein and the rho-GDP dissociation inhibitor from bovine neutrophil cytosol. Effect of phosphoinositides on rho ADP-ribosylation by the C3 exoenzyme of Clostridium botulinum. Biochemistry. 1992 Dec 29;31(51):12863–12869. [PubMed]
  • Sekine A, Fujiwara M, Narumiya S. Asparagine residue in the rho gene product is the modification site for botulinum ADP-ribosyltransferase. J Biol Chem. 1989 May 25;264(15):8602–8605. [PubMed]
  • Ueda T, Kikuchi A, Ohga N, Yamamoto J, Takai Y. Purification and characterization from bovine brain cytosol of a novel regulatory protein inhibiting the dissociation of GDP from and the subsequent binding of GTP to rhoB p20, a ras p21-like GTP-binding protein. J Biol Chem. 1990 Jun 5;265(16):9373–9380. [PubMed]
  • Paterson HF, Self AJ, Garrett MD, Just I, Aktories K, Hall A. Microinjection of recombinant p21rho induces rapid changes in cell morphology. J Cell Biol. 1990 Sep;111(3):1001–1007. [PMC free article] [PubMed]
  • Just I, Fritz G, Aktories K, Giry M, Popoff MR, Boquet P, Hegenbarth S, von Eichel-Streiber C. Clostridium difficile toxin B acts on the GTP-binding protein Rho. J Biol Chem. 1994 Apr 8;269(14):10706–10712. [PubMed]
  • von Eichel-Streiber C, Harperath U, Bosse D, Hadding U. Purification of two high molecular weight toxins of Clostridium difficile which are antigenically related. Microb Pathog. 1987 May;2(5):307–318. [PubMed]
  • Ohishi I, Iwasaki M, Sakaguchi G. Purification and characterization of two components of botulinum C2 toxin. Infect Immun. 1980 Dec;30(3):668–673. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Safer D. An electrophoretic procedure for detecting proteins that bind actin monomers. Anal Biochem. 1989 Apr;178(1):32–37. [PubMed]
  • O'Farrell PH. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PubMed]
  • Cooper JA. Effects of cytochalasin and phalloidin on actin. J Cell Biol. 1987 Oct;105(4):1473–1478. [PMC free article] [PubMed]
  • Narumiya S, Morii N. rho gene products, botulinum C3 exoenzyme and cell adhesion. Cell Signal. 1993 Jan;5(1):9–19. [PubMed]
  • Aktories K, Mohr C, Koch G. Clostridium botulinum C3 ADP-ribosyltransferase. Curr Top Microbiol Immunol. 1992;175:115–131. [PubMed]
  • Krautwurst D, Hescheler J, Arndts D, Lösel W, Hammer R, Schultz G. Novel potent inhibitor of receptor-activated nonselective cation currents in HL-60 cells. Mol Pharmacol. 1993 May;43(5):655–659. [PubMed]
  • Takai Y, Kaibuchi K, Kikuchi A, Kawata M. Small GTP-binding proteins. Int Rev Cytol. 1992;133:187–230. [PubMed]
  • Schiavo G, Benfenati F, Poulain B, Rossetto O, Polverino de Laureto P, DasGupta BR, Montecucco C. Tetanus and botulinum-B neurotoxins block neurotransmitter release by proteolytic cleavage of synaptobrevin. Nature. 1992 Oct 29;359(6398):832–835. [PubMed]
  • Popoff MR, Rubin EJ, Gill DM, Boquet P. Actin-specific ADP-ribosyltransferase produced by a Clostridium difficile strain. Infect Immun. 1988 Sep;56(9):2299–2306. [PMC free article] [PubMed]

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