Bacteriocin 28b production is induced by mitomycin in wild-type Serratia marcescens 2170 but not in Escherichia coli harboring the bacteriocin 28b structural gene (bss). Studies with a bss-lacZ transcriptional fusion showed that mitomycin increased the level of bss gene transcription in S. marcescens but not in the E. coli background. A S. marcescens Tn5 insertion mutant was obtained (S. marcescens 2170 reg::Tn5) whose bacteriocin 28b production and bss gene transcription were not increased by mitomycin treatment. Cloning and DNA sequencing of the mutated region showed that the Tn5 insertion was flanked by an SOS box sequence and three genes that are probably cotranscribed (regA, regB, and regC). These three genes had homology to phage holins, phage lysozymes, and the Ogr transcriptional activator of P2 and related bacteriophages, respectively. Recombinant plasmid containing this wild-type DNA region complemented the reg::Tn5 regulatory mutant. A transcriptional fusion between a 157-bp DNA fragment, containing the apparent SOS box upstream of the regA gene, and the cat gene showed increased chloramphenicol acetyltransferase activity upon mitomycin treatment. Upstream of the bss gene, a sequence similar to the consensus sequence proposed to bind Ogr protein was found, but no sequence similar to an SOS box was detected. Our results suggest that transcriptional induction of bacteriocin 28b upon mitomycin treatment is mediated by the regC gene whose own transcription would be LexA dependent.Serratia marcescens has been shown to produce bacteriocins upon induction with DNA-damaging agents (50). These bacteriocins have been classified into two groups (23): fraction 1 bacteriocins are active against Escherichia coli but not against S. marcescens, and fraction 2 bacteriocins are active against S. marcescens but not against E. coli (50). Bacteriocins belonging to fraction 1 are simple polypeptides that resemble colicins (50). Only colicin-like bacteriocins L and 28b have been studied in some detail. Bacteriocin L from S. marcescens JF246 has been isolated and characterized, and the effects of this bacteriocin on the incorporation of labelled leucine and thymidine and on the cellular levels of ATP in E. coli were similar to those produced by pore-forming colicins (17,18,40). On the other hand, the bacteriocin 28b structural gene (bss) has been cloned and sequenced, and the predicted amino acid sequence of the C-terminal part of this bacteriocin has been shown to have a high degree of similarity to the C-terminal domains of pore-forming colicins (56). The two bacteriocins are very closely related, if not the same, and similar bacteriocins are produced by most S. marcescens biotypes (21).Colicin production is induced by mitomycin and other DNAdamaging agents (38). Determinants for colicin production studied so far are encoded by either small high-copy-number colicinogenic plasmids (type I) or large low-copy-number colicinogenic plasmids (type II) (38). An important feature of these plasmids is that they confer on their host...
The gene encoding bacteriocin 28b from Serratia marcescens N28b (bss gene) has been cloned in Escherichia coli and its nucleotide sequence has been determined. The genetic determinants coding for other well-characterized bacteriocins from enterobacteria (colicins) are located in plasmids and they have always been shown to contain a gene responsible for immunity located downstream from the bacteriocin structural gene. In some cases there is another gene located downstream from the immunity gene, which is responsible for bacteriocin release. Analysis of bacteriocin 28b release and the sensitivity to this bacteriocin of E. coli strains harbouring recombinant plasmids containing the bss gene showed that bacteriocin 28b is not released from the cell in these strains and that their phenotypic insensitivity is not associated with any region close to the structural gene. The nucleotide sequence of the region downstream from the bss gene contains two putative open reading frames transcribed in the opposite direction to the bss gene. These open reading frames apparently encode proteins that seem not to be involved in bacteriocin immunity or release. Moreover, a S. marcescens N28b genomic library was screened and no immunity gene was found. Therefore, bacteriocin 28b differs greatly from the bacteriocins from other enterobacteria, and in the following senses it is unique: firstly, the gene encoding bacteriocin 28b seems to be located on the chromosome, and secondly, insensitivity to this bacteriocin in S. marcescens N28b is not associated with the expression of an immunity gene.
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