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Biomedical subjects

R J Rowbury

Publications and source records attributed to R J Rowbury.

At least 55 records · Page 3Linked to original sources
PubMed1987

Abortive infection by bacteriophage Me1 of Escherichia coli K12 strains bearing the plasmid ColV, I-K94.

Bacteriophage Me1 is unable to grow on Escherichia coli strains harbouring the ColV,I-K94 plasmid. The nature of this inhibition was investigated, and it was found not to be due to restriction, superinfection exclusion or receptor-mediated resistance, but to be a new example of plasmid-mediated abortive infection. Investigation of events occurring during abortive Me1 infection revealed some differences from previously described cases, especially with regard to late protein synthesis, which did occur, albeit showing abnormal amounts of some proteins. No major differences were observed in membrane permeability of productively and abortively infected cells. Phage-directed DNA synthesis was reduced in abortively infected cells. Comparative studies of Me1 and T4 revealed a striking similarity despite some minor differences.

Bacteriocin Plasmids↗
PubMed1987

Altered phage P1 attachment to strains of Escherichia coli carrying the plasmid ColV,I-K94.

Phages P1vir and P1cmclrf100 failed to form plaques on or multiply in Escherichia coli strains carrying the ColV,I-K94 plasmid; with P1cmclr100, the effect occurred both with phage from the lytic cycle and with that induced from a lysogen. The effect was on attachment, these P1 phages attaching poorly to ColV,I-K94+ strains. This receptor defect appeared to result mainly from the presence of ColV-encoded transfer and colicin components in the cells carrying ColV,I-K94 and it was specific to this plasmid. Phage Mu (which uses an attachment mechanism similar to that of phage P1) in the G(+) form attached to both Col- and ColV,I-K94+ strains but the G(-) form attached to neither type.

Bacteriocin Plasmids↗
PubMed1987

Increased sensitivity to erythromycin in Escherichia coli associated with the presence of the ColV,I-K94 virulence plasmid.

Introduction of the virulence plasmid, ColV,I-K94, into Escherichia coli strains led to increased sensitivity to erythromycin. This was the result of increased passage of antibiotic into ColV,I-K94+ organisms because the plasmid effect was abolished in bacteria which had been made permeable by chemical treatment. Full sensitivity in ColV+ strains appears to depend on the simultaneous presence of transfer and colicin components. Increased erythromycin sensitivity associated with the plasmid was demonstrated in organisms grown at 37 degrees C; the sensitivity of ColV,I-K94+ organisms grown at 25 degrees C was similar to that of the parent strain. Added Mg++ or Ca++ ions reversed erythromycin inhibition in strains with the basal level of sensitivity (i.e., the Col- parent grown at 25 degrees C or 37 degrees C or the ColV,I-K94+ derivative grown at 25 degrees C) and in those with the plasmid-associated increase in sensitivity. Addition of phosphate or EDTA to broth increased erythromycin sensitivity in Col- and ColV,I-K94+ strains although the latter was affected most. Erythromycin was more inhibitory at pH 8.5 than at pH 7.4. This enhanced activity was more marked against the ColV,I-K94+ strain than against the Col- strain. The effects of growth in phosphate-containing medium and at alkaline pH were partially additive. We suggest that ColV,I-K94+ strains may be sensitive to erythromycin because ColV-specified proteins are extruded by a process of "self-promoted transfer" and that the effects of these proteins on the lipopolysaccharide component of the outer membrane facilitates antibiotic influx.

Bacteriocin Plasmids↗
PubMed1987

Stimulatory effect of phosphate on colicin V synthesis/secretion by strains of Escherichia coli.

Larger inhibitory zones were produced by ColV+ strains on sensitive indicators if the strains were grown on nutrient agar and phosphate rather than nutrient agar. This was due to an effect of phosphate on colicin V synthesis/secretion and not to an effect on the response of the indicator. Phosphate produced an effect at 25 degrees C as well as at 37 degrees C (although zones were larger at 37 degrees C). It seems likely that phosphate stimulation of colicin V synthesis/secretion results from its effect in reducing divalent cation levels in NA because adding EDTA to NA also increased colicin V synthesis/secretion whereas the addition of magnesium or calcium salts had the opposite effect. It is possible that the extent of colicin V synthesis/secretion may depend on the strength of the LPS-LPS bonds in the outer membrane. Phosphate also stimulated synthesis/secretion of colicins E4, G and N but not of the other colicins or microcins tested. Studies with a wild Col+ E. coli isolate suggest that production of some colicins may not be detected on nutrient agar and that addition of phosphate may reveal the synthesis of new colicins.

Calcium↗
PubMed1986

Suppression by the ColV,I-K94 plasmid of a growth lesion in ompA mutants of Escherichia coli.

Organisms of three independently isolated ompA mutants of Escherichia coli failed to form colonies on glucose minimal agar (glucose MA) at 44 degrees C after growth in glucose minimal salts medium at 37 degrees C, although all three strains formed colonies on nutrient agar at 44 degrees C. Supplementation of the glucose MA with individual amino acids including L-methionine and/or L-cysteine did not allow colony formation at 44 degrees C, although addition of 0.1% Casamino acids was effective; replacement of glucose with other energy sources or ammonium ions with glutamate also did not allow growth at 44 degrees C. The failure to form colonies at 44 degrees C was not due to killing of the organisms, because colonies were formed if plates of the ompA mutant initially incubated at 44 degrees C were shifted to 30 degrees C after 16 h. Introduction of the ColV, I-K94 plasmid into P678-54 ompA, 1131 ompA or an ompC ompA mutant suppressed the 44 degrees C growth lesion, but other plasmids (F lac, R483ColIa, RI, ColB-K98, R124) tested in P678-54 ompA did not. Growth of the ColV, I-K94+ derivative at 44 degrees C was due to a suppressing effect of the plasmid rather than to introduction of the plasmid into a variant with normal or altered OmpA protein. An attempt was made to ascertain which component(s) encoded by ColV, I-K94 was (were) responsible for allowing growth at 44 degrees C. Transfer components appeared unlikely to be involved and plasmids which conferred individual colicins (plus the corresponding immunity component) did not suppress.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Outer Membrane Proteins↗
PubMed1986

Virulence plasmid-associated sensitivity to acid in Escherichia coli and its possible significance in human infections.

Several strains of Escherichia coli were markedly sensitised to killing at pH 2.5 or 3.5 when the ColV,I-K94 virulence plasmid was introduced into them. For strain 1829, the effect on acid sensitivity was due to the presence of plasmid in the previously resistant strain rather than to its introduction into an acid-sensitive variant already in the population. Acid sensitivity was also conferred by the ColV-K30 and ColB-K98 plasmids and the resistance plasmid R124-F2; other plasmids tested had no marked effect. Studies of ColV+ strains carrying mutant plasmids indicated that it was the presence of ColV-encoded transfer components that made ColV,I-K94+ strains acid-sensitive. Organisms in the exponential phase of growth were more sensitive to acid than were those from stationary phase cultures and this difference was more marked for ColV,I-K94+ strains than for Col- ones. Moreover, ColV+ strains exposed to conditions of low pH for short periods subsequently grew less well than the Col- parent and appeared to be sensitised by the damage to the effects of H2O2. These results indicate that some ColV+ strains may be more sensitive to gastric acid and to phagocytic acidity than are Col- strains. ColV,I-K94+ strains grew as well as Col- ones in broth or urine at pH 4.5-6.0 which suggests that the presence of the plasmid would not be detrimental to bacterial growth in the urinary tract. The presence of transfer components in the outer membrane of ColV,I-K94+ bacteria may destabilize the lipopolysaccharide layer allowing increased penetration of hydrogen ions.

Acids↗
PubMed1986

Virulence plasmid-associated adhesion of Escherichia coli and its significance for chlorine resistance.

Introduction of the ColV, I-K94 virulence plasmid into strains of Escherichia coli led (for four out of five strains tested) to a marked increase in the ability of organisms to adhere to glass beads. For strain 1829, the plasmid led to increased attachment to other materials including sand, agar, agarose, chitin and cellulose. The increased adhesion to glass beads was due to the presence of the plasmid and not to its introduction into a variant with altered adhesive properties. The plasmid-encoded VmpA protein did not appear to be necessary for the ColV, I-K94-promoted adhesion but adhesion was absolutely dependent on the presence of derepressed levels of transfer components in the ColV+ strains and partially dependent on the presence of colicin components. The extent of the plasmid-promoted adhesion was greatest for organisms grown at 30 degrees, 37 degrees or 42 decrees C and adhesion was almost abolished by growth at 21 degrees or 25 degrees C; this finding is in accord with transfer and colicin components being involved in adhesion. Of several other plasmids tested for their effects on adhesion, those with derepressed transfer properties showed a marked effect as did the RI resistance plasmid. Because of the ease of handling glass bead-attached organisms, such preparations were used as a model for studying the relevance of attachment to the resistance of E. coli to chlorination in the water purification process. Organisms of 1829 ColV, I-K94, attached to glass beads, were more resistant to damage and killing by chlorine than were unattached organisms. Three findings suggest that such chlorine resistance may be significant for survival during water chlorination. Firstly, ColV, I-K94+ bacteria became attached if incubated in sewage effluent with glass beads at 20 degrees C. Secondly, ColV+ organisms already attached to glass beads maintained their attachment during 24 h incubation in effluent at 20 degrees C and thirdly such effluent incubated organisms remained chlorine resistant provided that they retained their attachment.

Bacterial Adhesion↗
PubMed1985

Effects of the virulence plasmid ColV, I-K94 on the sensitivity of Escherichia coli to putative environmental inhibitory agents.

Derivatives of Escherichia coli carrying the virulence plasmid, ColV, I-K94 were more resistant than the ColV- parents to phage Mel but were more sensitive to the hydrophobic inhibitors deoxycholate, erythromycin and lysozyme. The basis for these changes in sensitivity has been examined in ColV+ mutants with altered colicin or VmpA protein levels and in ColV+ strains with repressed transfer properties.

Bacterial Outer Membrane Proteins↗
PubMed1985

The effects of the virulence plasmid ColV, I-K94 on the survival of Escherichia coli in sewage effluent.

Escherichia coli 1829 and its mucoid and non-mucoid ColV, I-K94+ derivatives (1829 ColVMuc and 1829 ColVNM) were tested for survival in final sewage effluent (FE) at 20 degrees C. All three strains survived well for 24 h; in some experiments there was also good survival at 48 h but in others there was marked loss of viability. There were only small differences between the strains in their ability to survive in FE i.e. neither ColV, I-K94 nor the mucoid character consistently increased or reduced survival markedly. To assess which factors in FE influence survival, the three strains were also tested in heated FE, filtered FE and chloroform-treated FE. All three strains survived well in heated FE at 20 degrees C but 1829 ColVNM, lost viability much more markedly than the other two strains in filtered FE or chloroform-treated FE. In spite of the poor nutrient level in FE, there was a marked increase, in bacteriophage number in those cultures incubated in chloroform treated FE.

Bacteriocin Plasmids↗
PubMed1984

Effects of the resistance plasmid R124 on the level of the OmpF outer membrane protein and on the response of Escherichia coli to environmental agents.

The introduction of the F-like resistance plasmid R124 into an ompC mutant of Escherichia coli K12 conferred altered sensitivity to a wide range of inhibitory agents. Sensitivity to ampicillin, chloramphenicol, ethionine, copper ions, deoxycholate, two fatty acids and colicins L and M was decreased by the plasmid. In contrast the plasmid-bearing ompC derivatives were more sensitive than the plasmid-free ompC mutant to erythromycin, cetyltrimethylammonium bromide and phenol. Introduction of R124 into the ompC strain also decreased the level of the OmpF protein and some (but not all) of the changed sensitivities listed above clearly resulted from this outer membrane protein deficiency. The presence in the ompC mutant of R124 (rather than the more efficient introduction of the plasmid into variants of the ompC strain) led to at least most of the changes described above because those tested were accentuated by the presence of a copy mutant of R124 and reversed by plasmid curing.

Cephaloridine↗
PubMed1982

A new major outer membrane protein in derivatives of Escherichia coli carrying the virulence plasmid ColV-K94.

Strains of Escherichia coli K12 harbouring ColV-K94 contain a new major outer membrane protein of molecular weight ca. 33,000. The new protein resembles the Omp A protein in that (1) it is trypsin-sensitive in membrane preparations and (2) it is not murein-associated. The new protein cannot, however, replace the Omp A protein as a functional receptor for the phages K3 and TuII*, for colicin L or for efficient conjugation with F-like donors. The new protein is apparently not a transfer component and is produced by cells unable to produce colicin. Labelling experiments with minicells suggest that the new protein is one of two major plasmid-encoded membrane proteins.

Bacterial Outer Membrane Proteins↗
PubMed1980

Host components required for the replication of the resistance plasmid R124 and a copy mutant derivative.

The replication of R124, and a copy mutant derivative of it, was measured with respect to dependence on the host DnaA, DnaB, DnaC, DnaE, DnaG, and PolA gene products. Both plasmids replicated under conditions where the DnaA gene product was inactivated or where the polymerising activity of the PolA gene product was reduced. In contrast, neither plasmid replicated to any appreciable extent, if the DnaB, DnaC, DnaE or DnaG gene products were inactivated. R124 integratively suppressed the lesion of the dnaA mutant but the copy mutant derivative had only a very weak suppressing effect. Neither plasmid suppressed the lesions of any of the other dna mutants.

Bacterial Proteins↗
PubMed1979

Alterations in the cell envelope composition of Proteus mirabilis during the development of swarmer cells.

Long, swarming cells of Proteus mirabilis had different proportions of some lipopolysaccharide components when compared to short cells, either agar grown or broth grown. Fluorescence spectrophotometry of antibody binding, and sodium dodecyl sulphate-polyacrylamide gel electrophoresis indicated that the change was in the proportion of lipopolysaccharide with long O-antigenic sidechains, swarmer lipopolysaccharide relative to short sidechain lipopolysaccharide than the non-swarming cells. The proteins and phospholipids of the envelop remained the same during swarmer development. The results are discussed in relation to the increase in flagella synthesis and permeability to some antibacterial agents during swarmer development.

Binding Sites, Antibody↗
PubMed1979

Factors influencing the copy number of F-like plasmids in Escherichia coli and Salmonella typhimurium.

The copy numbers of Flac, four F-like plasmids and pLT2 were estimated in two strains of Salmonella typhimurium and (for all except pLT2) one strain of Escherichia coli. For organisms grown in casamino acids minimal medium, the plasmids spanned a 7--8 fold range of copy number with ColB-K98 having the highest copy number in each strain and R124 the lowest. The copy number of ColB-K98 was substantially greater than 1 in each of the strains tested. There was no clear relation between the plasmid size and copy number, although the plasmids studied spanned only a narrow size range. The copy number of individual plasmids was slightly reduced or not affected at all by the presence of a second plasmid in the same strain. Derivatives harbouring each of the plasmids were grown in three different media to ascertain how plasmid copy number responds to changes in growth rate. For each plasmid, the copy number increased with decreasing growth rate. Extracts from each of the three strains harbouring ColB-K98 contained two distinct plasmid species. One appeared to be about twice as large as the other and both were absent from Col- segregants.

Conjugation, Genetic↗
PubMed1978

An alteration in outer membrane permeability associated with a division lesion in a strain of Salmonella typhimurium.

Salmonella typhimurium strain 4a is a temperature sensitive mutant with defects in both septation and separation. The separation lesion was reversed by phenethylalcohol but this agent failed to allow septation or growth at restrictive temperature. Organisms of strain 4a grown at 42 degrees C were, unlike the parental strain, resistant to lysis by lysozyme plus EDTA and lipopolysaccharide was poorly extracted by EDTA from cultures of strain 4a grown at 42 degrees C. Such cultures may, therefore, be resistant to lysis with lysozyme plus EDTA not because the murein is altered but because the EDTA fails to permeabilize the outer membrane to lysozyme. In confirmation of this, murein isolated from strain 4a after growth at 42 degrees C showed the same sensitivity to lysozyme as murein from the parental strain. In spite of the altered envelope properties of strain 4a after growth at 42 degrees C, no major changes in protein or phospholipid composition have so far been demonstrated.

Cell Division↗
PubMed1977

A mutant of the antibiotic resistance factor R124 with altered copy number.

A mutation conferring increased antibiotic resistance on Salmonella typhimurium strain 11G carrying R124 was plasmid determined; strains harbouring the mutant plasmid contained more DNA as ccc plasmid than those harbouring R124. The increased copy number was manifested at all growth rates tested.

Anti-Bacterial Agents↗
PubMed1977

An effect of F-like plasmids on the maintenance of Flac in a dnaC mutant of Salmonella typhimurium.

Flac maintenance was aberrant at permissive temperature in a temperature-sensitive dnaC mutant of Salmonella typhimurium when the normally resident pLT2 plasmid was present. Flac was, however, efficiently transferred into the dnaC pLT2+ strain and the resulting Flac derivative was almost as efficient in transferring Flac as was dnaC+ pLT2+ Flac strains indicating that aberrant Flac maintenance was not associated with appreciable inhibition of transfer replication. A range of F-like plasmids behaved like pLT2 in causing aberrant Flac maintenance when present in the dnaC pLT2- strain. Flac was, however, stably maintained in the dnaC strain in the absence of other plasmids. Although the F-like plasmids destabilized Flac, each was stably maintained when introduced into strain 11G dnaC pLT2+ and pLT2 was also apparently stable under these conditions. The destabilizing effect of pLT2 and other fi+ plasmids was not consequent upon their inhibiting the formation of a repressible F transfer component needed for Flac replication in the dnaC strain. Incompatibility between Flac and the other plasmids induced by the dnaC lesion also appeared unlikely to be a cause of the aberrant Flac maintenance. The possibility is discussed that the initiation of Flac replication differs from that of pLT2 and the F-like plasmids with F competing less effectively than the others for the DnaC gene product.

Conjugation, Genetic↗