Cloning and expression of the dermonecrotic toxin gene of Pasteurella multocida ssp. multocida in Echerichia coli

Kamps, A. M. I. E.; Kamp, E. M.; Smits, M.A.

doi:10.1111/j.1574-6968.1990.tb13860.x

Cited by 23 publications

(8 citation statements)

References 6 publications

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“…Once it was discovered that only certain capsular serotype D and A strains of P. multocida were responsible for chronic turbinate atrophy, a large protein toxin was identified, isolated, and cloned from these strains (148,194,196,461,462). The purified, 146-kDa P. multocida toxin (PMT), encoded by the toxA gene located on a putative lysogenic bacteriophage (463), was subsequently demonstrated to be the primary agent responsible for the symptoms of atrophic rhinitis (461,(464)(465)(466)(467)(468)(469)(470)(471), as well as a number of other clinical symptoms associated with P. multocida infection (189,190,192,461,464,470,(472)(473)(474)(475). PMT-mediated bone atrophy appears to occur through disruption of normal cell signaling processes in bone-generating osteoblasts and macrophage-like osteoclasts (205,206,466,(476)(477)(478)(479)(480)(481)(482).…”

Section: Survival In the Host Environmentmentioning

confidence: 99%

Pasteurella multocida: from Zoonosis to Cellular Microbiology

2013

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SUMMARY In a world where most emerging and reemerging infectious diseases are zoonotic in nature and our contacts with both domestic and wild animals abound, there is growing awareness of the potential for human acquisition of animal diseases. Like other Pasteurellaceae , Pasteurella species are highly prevalent among animal populations, where they are often found as part of the normal microbiota of the oral, nasopharyngeal, and upper respiratory tracts. Many Pasteurella species are opportunistic pathogens that can cause endemic disease and are associated increasingly with epizootic outbreaks. Zoonotic transmission to humans usually occurs through animal bites or contact with nasal secretions, with P. multocida being the most prevalent isolate observed in human infections. Here we review recent comparative genomics and molecular pathogenesis studies that have advanced our understanding of the multiple virulence mechanisms employed by Pasteurella species to establish acute and chronic infections. We also summarize efforts being explored to enhance our ability to rapidly and accurately identify and distinguish among clinical isolates and to control pasteurellosis by improved development of new vaccines and treatment regimens.

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Section: Survival In the Host Environmentmentioning

confidence: 99%

Pasteurella multocida: from Zoonosis to Cellular Microbiology

2013

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“…Pasteurella multocida toxin (PMT) has been shown to be an extremely potent and effective mitogen for fibroblast cell lines and early-passage cultures (4). The toxin is a monomeric 146-kDa protein that has been purified (5)(6)(7)(8), cloned (9)(10)(11), and sequenced (12)(13)(14). The deduced amino acid sequence of PMT did not reveal any significant homologies with other toxins or proteins (13,14).…”

mentioning

confidence: 99%

Pasteurella multocida toxin is a potent inducer of anchorage-independent cell growth.

Higgins

Murphy

Staddon

et al. 1992

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

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The growth of many normal cells requires contact with an adhesive substratum, a requirement that is frequently abrogated in the transformed phenotype. We have explored pathways that can lead to the anchorage-independent growth of cultured Rat-i fibroblasts. PasteureUa mulocida toxin (PMT), a 146-kDa mitogenic protein, caused a striking increase in the formation of colonies (>200 jam) from single cells in soft agar. The magnitude of the effect of PMT was greater than that achieved by epidermal growth factor or platelet-derived growth factor. The toxin was extremely potent, with half-maximal and maximal effects observed at 1 and 10 pM PMT, respectively. This concentration dependence of the action of the toxin is similar to that for the stimulation of DNA synthesis in adherent cultures of the cells. Stimulation ofcolony formation could be achieved by a transient exposure of the cells to PMT and it was blocked by methylamine, indicating that the toxin enters the cells to act. Colony formation was stimulated equally by native and recombinant PMT, but a truncated version (33.5 kDa) of the recombinant toxin was ineffective. PMT antiserum blocked colony formation in response to PMT. In the Rat-1 cells, PMT stimulated the phospholipase C-mediated hydrolysis of inositolphospholipids, as indicated by the stimulation of inositol phosphate release, Ca2+ mobilization, and phosphorylation of a protein kinase C substrate. The results indicate that the deregulation of signal-transduction pathways as elicited by an intracellularly acting bacterial toxin can induce a malignant phenotype.The mechanisms of action of bacterial toxins have provided insights into the control of cellular regulatory processes, including signal transduction and cell proliferation (1-3). Pasteurella multocida toxin (PMT) has been shown to be an extremely potent and effective mitogen for fibroblast cell lines and early-passage cultures (4). The toxin is a monomeric 146-kDa protein that has been purified (5-8), cloned (9-11), and sequenced (12-14). The deduced amino acid sequence of PMT did not reveal any significant homologies with other toxins or proteins (13,14). Both native and recombinant PMT (rPMT) are mitogenic at picomolar concentrations (4). Several lines of evidence indicate that PMT enters the cells and acts intracellularly to initiate and sustain DNA synthesis. Thus, a transient exposure of the cells to the toxin was sufficient to commit them to S phase and cell division. Furthermore, early but not late addition of the lysosomotrophic agent methylamine selectively blocked the mitogenic action of rPMT (4).Prior to the stimulation of DNA synthesis, rPMT enhanced the formation of inositol phosphates (15), increased the cellular content of diacylglycerol, caused the translocation of protein kinase C, and stimulated the phosphorylation of an 80-kDa protein (16) that is a major substrate of protein kinase C (refs. 17-19, and see ref. 20). Furthermore, the binding of epidermal growth factor (EGF) to its receptor was decreased by rPMT, an action ...

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“…The toxin is also mitogenic for the following established cell lines, Rat-1 [3], BALB/c 3T3, NIH 3T3, 3T6 and human fibroblasts [1]. 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].…”

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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Cloning and expression of the dermonecrotic toxin gene of Pasteurella multocida ssp. multocida in Echerichia coli

Cited by 23 publications

References 6 publications

Pasteurella multocida: from Zoonosis to Cellular Microbiology

Pasteurella multocida: from Zoonosis to Cellular Microbiology

Pasteurella multocida toxin is a potent inducer of anchorage-independent cell growth.

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

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