Cytotoxic necrotizing factor type 2 produced by virulent Escherichia coli modifies the small GTP-binding proteins Rho involved in assembly of actin stress fibers.

Oswald, Éric; Sugai, Motoyuki; Labigne, Agnès; Wu, Henry C.; Fiorentini, Carla; Boquet, Patrice; O'Brien, A D

doi:10.1073/pnas.91.9.3814

Cited by 178 publications

(178 citation statements)

References 41 publications

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“…Several lines of evidence indicate that the N-terminal region of the toxin binds to target cells and that the C-terminal region carries the intracellularly active moiety, the nature of which remains unknown (15)(16)(17)(18). The N-terminal region is partly homologous to Escherichia coli cytotoxic necrotizing factors, CNF1 and CNF2 (19,20). In contrast, in the C-terminal region of PMT, there is neither a known catalytic motif nor a region homologous with any other toxin or enzyme, and therefore no structural information has been available for elucidation of the intracellular action of the toxin.…”

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confidence: 99%

Crystal structures reveal a thiol protease-like catalytic triad in the C-terminal region ofPasteurella multocidatoxin

Kitadokoro

Kamitani²,

Miyazawa³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

134

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Pasteurella multocida toxin (PMT), one of the virulence factors produced by the bacteria, exerts its toxicity by up-regulating various signaling cascades downstream of the heterotrimeric GTPases Gq and G12/13 in an unknown fashion. Here, we present the crystal structure of the C-terminal region (residues 575-1,285) of PMT, which carries an intracellularly active moiety. The overall structure of C-terminal region of PMT displays a Trojan horse-like shape, composed of three domains with a ''feet''-,''body''-, and ''head''-type arrangement, which were designated C1, C2, and C3 from the N to the C terminus, respectively. The C1 domain, showing marked similarity in steric structure to the N-terminal domain of Clostridium difficile toxin B, was found to lead the toxin molecule to the plasma membrane. The C3 domain possesses the Cys-HisAsp catalytic triad that is organized only when the Cys is released from a disulfide bond. The steric alignment of the triad corresponded well to that of papain or other enzymes carrying CysHis-Asp. PMT toxicities on target cells were completely abrogated when one of the amino acids constituting the triad was mutated. Our results indicate that PMT is an enzyme toxin carrying the cysteine protease-like catalytic triad dependent on the redox state and functions on the cytoplasmic face of the plasma membrane of target cells.crystallography ͉ cysteine protease ͉ membrane targeting

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mentioning

confidence: 99%

Crystal structures reveal a thiol protease-like catalytic triad in the C-terminal region ofPasteurella multocidatoxin

Kitadokoro

Kamitani²,

Miyazawa³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

134

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“…The cnf2 gene is indeed carried by the Vir plasmid (Oswald et al, 1994a), whereas the cnf1 gene is located on the chromosome (Falbo et al, 1992).…”

Section: Introductionmentioning

confidence: 99%

“…NTEC2 strains have been mainly isolated from ruminants with intestinal infections or with septicemia Cid et al, 1996;Pohl et al, 1997). The necrotoxigenic E. coli produce two different toxins (CNF1 and CNF2) closely related antigenically and genetically (De Rycke et al, 1987;Oswald et al, 1994a).…”

Section: Introductionmentioning

confidence: 99%

Characteristics of necrotoxigenic Escherichia coli isolated from septicemic and diarrheic calves between 1958 and 1970

Bost

Bâbe

Jacquemin

et al. 2001

Veterinary Microbiology

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A total of 434 Escherichia coli isolated from septicemic calves between 1958 and 1965 and 430 E. coli isolated from diarrheic calves between 1967 and 1970 were studied by colony hybridisation and PCR assays for the presence of the cnf1-and the cnf2-like genes. They were also studied for the presence of genes coding for putative virulence factors associated with the CNF toxins including F17-, Pap-and Sfa-®mbrial adhesins and the recently described CDT-III toxin and AfaVIII-a®mbrial adhesin. Thirty (7%) of the 434 septicemic strains were positive for CNF by colony hybridisation. Twenty-six were con®rmed as necrotoxigenic E. coli type 2 (NTEC2) and four as NTEC1 by PCR. Thirty-®ve (8%) of the 430 diarrheic strains were positive for CNF by colony hybridisation. Five of them were studied by PCR and con®rmed as NTEC1. The 26 septicemic NTEC2 strains and 20 of the 35 diarrheic NTEC including three of the ®ve NTEC1 were positive for CDT-III. All adhesins studied were present in NTEC as well as in non-NTEC. NTEC1 were mainly Pap-, Sfa-and/or Afa8-positive, whereas NTEC2 were mainly F17-and/or Afa8-positive. This study shows that necrotoxigenic E. coli with their associated adhesins and toxins were present in calves as early as 1958, but their prevalence seems to have increased since that time. #

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“…The toxin has been cloned, sequenced and expressed in Escherichia co# [4][5][6][7][8][9]. PMT is encoded as a 146 kDa singlechain bacterial protein, which shares partial homology with the multinucleating toxins, cytotoxic necrotising factors types 1 and 2 (CNF-1, CNF-2), produced by some strains of E. coli [10,11]. Although PMT contains a putative ADP-ribosylation motif [6], recent data have shown that PMT is unlikely to ADP ribosylate its substrate [12,13].…”

Section: Introductionmentioning

confidence: 99%

The potent mitogen Pasteurella multocida toxin is highly resistant to proteolysis but becomes susceptible at lysosomal pH

Smyth¹,

Pickersgill²,

Lax³

1995

FEBS Letters

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The susceptibility of the potent mitogen Pasteurella multocida toxin (PMT) to various proteases was investigated. PMT at a toxin to protease molar ratio of 1 : 1 was resistant to 8 of the 11 proteases tested after one hour. With longer incubation, PMT remained resistant to 7 proteases, and this correlated with a retention of biological activity, indicating that PMT might not require proteolytic cleavage at least until it bound to a cell receptor. Previous evidence had suggested that PMT is processed in the cell via an endosome or lysosome. We have shown that PMT became susceptible to proteolysis when the pH was lowered to 5 or below. This supports the previous suggestion that PMT is processed via a low pH compartment in the cell.

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Cytotoxic necrotizing factor type 2 produced by virulent Escherichia coli modifies the small GTP-binding proteins Rho involved in assembly of actin stress fibers.

Cited by 178 publications

References 41 publications

Crystal structures reveal a thiol protease-like catalytic triad in the C-terminal region ofPasteurella multocidatoxin

Crystal structures reveal a thiol protease-like catalytic triad in the C-terminal region ofPasteurella multocidatoxin

Characteristics of necrotoxigenic Escherichia coli isolated from septicemic and diarrheic calves between 1958 and 1970

The potent mitogen Pasteurella multocida toxin is highly resistant to proteolysis but becomes susceptible at lysosomal pH

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