Pseudomonas Aeruginosa Secretes and Correctly Processes Human Growth Hormone

Gray, Gregory L.; McKeown, Kathleen A.; Jones, Andrew J. S.; Seeburg, Peter H.; Heyneker, Herbert L.

doi:10.1038/nbt0284-161

Cited by 36 publications

(6 citation statements)

References 25 publications

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“…Each lane contained the material from two spots (1.5 x 108 cells) harvested from the TPMP plates. Protein bands were transferred to nitrocellulose as described previously (7). Filters were blocked and reacted with a mixture of rabbit anti-p-galactosidase and rabbit anti-protein S sera by using conditions described previously (16).…”

Section: Methodsmentioning

confidence: 99%

Control of developmental gene expression by cell-to-cell interactions in Myxococcus xanthus

Gill¹,

Cull²

1986

J Bacteriol

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The ssbA mutants of Myxococcus xanthus behave as if they are unable to produce a cell-to-cell signal required for normal development. They are unable to form fruiting bodies or spores on developmental medium. They do sporulate, however, if allowed to develop in mixtures with wild-type cells. Fusions of developmentally induced promoters of M. xanthus to the Escherichia coil lacZ gene were used to characterize the effect of the ssbA mutations on developmental gene expression. Each of the five independent fusions tested was found to be dependent upon the ssbA+ allele for full expression. The ssbA mutants were able to express each of these fusions if the mutants were allowed to develop in mixtures with wild-type (Lac-) cells. These results cannot be explained on the basis of genetic exchange. The data are consistent with regulation of gene expression mediated by cell-to-cell interactions.

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Section: Methodsmentioning

confidence: 99%

Control of developmental gene expression by cell-to-cell interactions in Myxococcus xanthus

Gill¹,

Cull²

1986

J Bacteriol

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“…For transformation with plasmids derived from pUC8 or pUC9 (12), E. coli strain JM83 (12) was used. pHGH207-1* is a plasmid that has been used for the expression of human growth hormone under E. coli trp promoter control (11). The M13 vectors mp8 and mp9 were used for all DNA sequence analyses as described (13).…”

Section: Methodsmentioning

confidence: 99%

“…E. coli 294 (11) was used for transformations with pBR322-derived plasmids. For transformation with plasmids derived from pUC8 or pUC9 (12), E. coli strain JM83 (12) was used.…”

Section: Methodsmentioning

confidence: 99%

“…complementary to the region of lowest genetic degeneracy (positions [8][9][10][11][12] were synthesized (15) and 32P-labeled at the 5' end with T4 polynucleotide kinase and [y-32P]ATP. The probe pool was hybridized to a recombinant pUC9-PA103 library of BamHI-cleaved genomic DNA constructed essentially as described (16).…”

Section: Tide Probes D(g-c-_-c-a-and-t-c-g-t-t-c-c-a) Potentiallymentioning

confidence: 99%

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Cloning, nucleotide sequence, and expression in Escherichia coli of the exotoxin A structural gene of Pseudomonas aeruginosa.

Gray¹,

Smith²,

Baldridge³

et al. 1984

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

250

118

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“…The transport of proteins through the inner membrane of bacteria has been extensively studied in E. coli (for recent reviews, see references 33,36,41) but only recently in P. aeruginosa (15,26,32,48). The latter is unusual among gram-negative organisms in being able to release a number of proteins into the medium during growth, while in the more typical E. coli, secreted proteins are almost always retained within the periplasm or the outer membrane.…”

mentioning

confidence: 99%

Nucleotide sequence and expression of a phosphate-regulated gene encoding a secreted hemolysin of Pseudomonas aeruginosa

Pritchard¹,

Vasil²

1986

J Bacteriol

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A 3.3-kilobase-pair fragment of Pseudomonas aeruginosa DNA containing the phospholipase C (heat-labile hemolysin) gene was sequenced, and the location of the gene was determined. The gene product contains at its NH2 terminus a 38-amino acid sequence which structurally resembles the signal peptides of other secreted proteins but is unusually long and positively charged (6+). The location of the translation start codon was determined by constructing a series of plasmids in which the promoter of a transcription vector was ligated to Pseudomonas DNA containing deletions at the 5' end of the gene. The plasmids were used to transform Escherichia coli, and the resulting clones were assayed for hemolysin activity. In addition, sizes of truncated proteins produced by mutants with translation terminators introduced at specific sites were analyzed in E. coli maxicells. The gene is transcribed, starting just upstream of the hemolysin gene, as an mRNA of approximately 2,800 bases. Analysis of the nucleotide sequence, analysis of mutants in maxicells, and transcriptional studies indicate that the hemolysin is part of an operon composed of two genes. Phosphate regulation of the operon is at the transcriptional level. The location of the 5' end of the transcript was determined by Si mapping.Phospholipase C (PLC) (phosphatidylcholine cholinephosphohydrolase) from Pseudomonas aeruginosa has been studied from different viewpoints: (i) as a hemolytic toxin that may contribute to the virulence of an opportunistic pathogen (22,23,44), (ii) as a secreted protein (10,12,25,48), and (iii) as a part of the phosphate regulon (13,14,19). Extracellular PLC hemolysins are also produced by organisms such as Clostridium perfringens (alpha toxin) and Staphylococcus aureus (beta toxin). In contrast, there is a different class of bacterial hemolysins produced by S. aureus (alpha toxin), Escherichia coli, and other organisms which are thought to have a nonenzymatic mechanism of action (18).The mechanism of pathogenicity for P. aeruginosa is complex, and the role of the hemolysin is not quite clear. PLC has been implicated as a virulence determinant in the pathogenesis of lung infection (22), and in another study, PLC production by urinary tract isolates was greater than that by lung, blood, or other isolates (4). Liu (23) observed that the PLC hemolysin and alkaline phosphatase are produced together during growth in low Pi and repressed during growth in high P, and proposed that these enzymes function cooperatively as a phosphate-scavenging mechanism. P. aeruginosa plcA mutants have been isolated which are deficient in the production of PLC and several other exported phosphate-repressible proteins (13, 17). Another class of mutants (plcB) hyperproduce most phosphaterepressible proteins and are also altered in Pi transport (14,17).PLC is also interesting for its properties as a secreted protein. The transport of proteins through the inner membrane of bacteria has been extensively studied in E. coli (for recent reviews, see references 33, 36, 41) bu...

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Pseudomonas Aeruginosa Secretes and Correctly Processes Human Growth Hormone

Cited by 36 publications

References 25 publications

Control of developmental gene expression by cell-to-cell interactions in Myxococcus xanthus

Control of developmental gene expression by cell-to-cell interactions in Myxococcus xanthus

Cloning, nucleotide sequence, and expression in Escherichia coli of the exotoxin A structural gene of Pseudomonas aeruginosa.

Nucleotide sequence and expression of a phosphate-regulated gene encoding a secreted hemolysin of Pseudomonas aeruginosa

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