The Regulation of Colicin Synthesis and Colicin Factor Transfer in Escherichia coli K 12
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Summary: Cells of Escherichia coli, newly infected with the colicin I factor (colI), showed an enhanced efficiency of transfer of this factor (HFC), and were also more likely to undergo lethal colicin synthesis, than were stably colicinogenic cells. Up to 20% of the cells of stably colI + strains were induced to produce colicin by ultraviolet irradiation, and from such irradiated cultures transfer of the colI factor occurred more efficiently. To account for these results, it is proposed that the colI factor exists as an autonomous non-chromosomal genetic element which sets up its own system of self-regulation within cells of stably colicinogenic strains.Keywords:
Colicin
A survey of colicins in the ECOR reference collection of Escherichia coli is presented. Twenty-five of the 72 ECOR strains exhibited a phenotype consistent with colicin production and E. coli isolated from human hosts were more likely to be colicinogenic than those from animal hosts. Multiple representatives of two Col plasmids, lowmolecular-mass ColE1 plasmids and high-molecular-mass, conjugative ColIa plasmids were isolated from the ECOR collection and were examined with a combination of restriction fragment and Southern analysis. These data suggested that ColE1 plasmids comprise a stable (cohesive) plasmid lineage, while ColIa plasmids represent a family of distinct plasmid lineages united by the presence of the colicin Ia operon.
Colicin
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The further identification of regions of the colicin E1 plasmid that affect plasmid functions has been achieved by studying deletions and TnA insertions of the plasmid. Colicin production, colicin immunity, relaxation of plasmid deoxyribonucleic acid, and plasmid incompatibility functions have been examined. A strong correlation has been observed between the ability of colicin E1 plasmid deoxyribonucleic acid to be relaxed and the ability of that plasmid to be transferred by conjugation.
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Escherichia coli infections can be a major health burden, especially with the organism becoming increasingly resistant to “last-resort” antibiotics such as carbapenems. Although colicins are potent narrow-spectrum antimicrobials with potential as future antibiotics, high levels of naturally occurring colicin insensitivity have been documented which could limit their efficacy. We identify O-antigen-dependent colicin insensitivity in a clinically relevant uropathogenic E. coli strain and show that this insensitivity can be circumvented by minor changes to growth conditions. The results of our study suggest that colicin insensitivity among E. coli organisms has been greatly overestimated, and as a consequence, colicins could in fact be effective species-specific antimicrobials targeting pathogenic E. coli such as uropathogenic E. coli (UPEC).
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A series of transformants have been derived which carry the Col plasmids from 11 E-colicin-producing strains isolated by Fredericq or Hamon. The ColE plasmids identified included four of type E1, five of type E2, and two of a type designated E4 by Horak (Zentralbl. Bakteriol. Parasitenkd. Infektionskr. Hyg. Abt. 1 Orig. Reihe A 233:58-63, 1975). Strain K317 was shown to carry a ColE7 plasmid and a 2.7-megadalton (Md) ColE2imm plasmid which confers immunity to colicin E2 but not the ability to produce colicin. Other plasmids identified in the colicinogenic isolates were a 3.4-Md ColN plasmid and a 1.3-Md Col plasmid of unknown type. The ColE1 plasmids all continued replicating in cultures treated with chloramphenicol. Strains carrying the ColE4 or ColE3-CA38 plasmid exhibited partial sensitivity to their own colicins and exhibited an unusual clearing-zone morphology when overlaid on stabs of colicin E2- or E7-producing strains. Except for ColE1-K53, ColE1-K47, ColE2-CA42, and ColE7-K317, all of the ColE plasmids were found to be about 4.3 Md in size. ColE2-CA42 and ColE7-K317 are both about 3.9 Md and are the only two plasmids of the E2, E3, E4, and E7 types that did not yield a 0.39-Md deoxyribonucleic acid fragment as an EcoRI digestion product. The comparisons of the ColE plasmids suggest several structural and functional relationships among them.
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To assess a collection of 96 Escherichia coli O157:H7 strains for their resistance potential against a set of colicinogenic E. coli developed as a probiotic for use in cattle.Escherichia coli O157:H7 strains were screened for colicin production, types of colicins produced, presence of colicin resistance and potential for resistance development. Thirteen of 14 previously characterized colicinogenic E. coli strains were able to inhibit 74 serotype O157:H7 strains. Thirteen E. coli O157:H7 strains were found to be colicinogenic and 11 had colicin D genes. PCR products for colicins B, E-type, Ia/Ib and M were also detected. During in vitro experiments, the ability to develop colicin resistance against single-colicin producing E. coli strains was observed, but rarely against multiple-colicinogenic strains. The ability of serotype O157:H7 strains to acquire colicin plasmids or resistance was not observed during a cattle experiment.Escherichia coli O157:H7 has the potential to develop single-colicin resistance, but simultaneous resistance against multiple colicins appears to be unlikely. Colicin D is the predominant colicin produced by colicinogenic E. coli O157:H7 strains.The potential for resistance development against colicin-based strategies for E. coli O157:H7 control may be very limited if more than one colicin type is used.
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Summary: We have tested the ability of pairs of colicin E plasmids to replicate stably in the same cell line. Although many of the pairs of E colicin plasmids were compatible, plasmids ColE3-CA38, ColE7-K317 and ColE8-J were mutually incompatible, as were ColE5-099, ColE6-CT14 and ColE9-J. Incompatibility between ColE6-CT14 and ColE5-099 or ColE9-J was asymmetrical, whereas incompatibility between the other plasmid pairs was symmetrical.
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The universal colicin-indicator strain Escherichia coli phi, unlike E. coli strain K-12, is sensitive to pesticin, a bacteriocin produced by wild-type Yersinia pestis. Eleven distinct phenotypes of E. coli phi mutants were obtained by selection for insensitivity to pesticin, group B colicins, the group A colicin S4, or coliphage T5. Representative isolates from eight of these classes closely resembled resistant receptor mutants (Cir-, Fep-, and TonA-) or tolerant mutants (TonB-, ExbB-, ExbC-, Ivt-, and Cmt-) described in Escherichia coli K-12. The remainder were unique; of these, one resembled TonB- but was also tolerant to colicin S4 (TonB/S4-), and the others exhibited specific resistance to either colicin S4 (Sfr-) or to pesticin (Psr-). All receptor mutants except Psr- remained sensitive to pesticin, whereas TonB/S4, TonB-, ExbB-, and ExbC- isolates were highly tolerant to this bacteriocin.
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ColE1
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Summary: Cells of Escherichia coli, newly infected with the colicin I factor (colI), showed an enhanced efficiency of transfer of this factor (HFC), and were also more likely to undergo lethal colicin synthesis, than were stably colicinogenic cells. Up to 20% of the cells of stably colI + strains were induced to produce colicin by ultraviolet irradiation, and from such irradiated cultures transfer of the colI factor occurred more efficiently. To account for these results, it is proposed that the colI factor exists as an autonomous non-chromosomal genetic element which sets up its own system of self-regulation within cells of stably colicinogenic strains.
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A collection of strains derived from Escherichia coli K12 W3110 and harbouring various colicin or microcin plasmids (18 and 2 representatives, respectively), or carrying well-characterized mutations conferring reduced colicin/microcin sensitivity is described. The strains can be used in typing schemes based on the identification of colicins, in the detection of new types of colicins/microcins, and in the further characterization of previously identified colicins/microcins and their plasmids.
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