The genetic switch for the regulatory pathway of Lactobacillus plantarum phage φg1e: characterization of the promoter PL, the repressor gene cpg, and the cpg-encoded protein Cpg in Escherichia coli

Kakikawa, Makiko; Ohkubo, Shingo; Syama, Michio; Taketo, Akira; Kodaira, Ken-Ichi

doi:10.1016/s0378-1119(99)00531-4

Cited by 18 publications

(12 citation statements)

References 27 publications

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“…The amino acid sequence of protein LA0195 is similar to that of the prophage repressor CI protein, and it contains a conserved DNA-binding domain. It is known that CI repressor can bind the specific DNA sequence upstream of and proximal to its own structure gene (7,9,34). Protein LA0195 was overexpressed and purified.…”

Section: Resultsmentioning

confidence: 99%

“…The EMSA results showed that protein LA0195 may bind to the strong promoter (tested in vivo) of transcripts I and II, although both of them lack the 14-bp dyad symmetry operator of CI protein binding sites (7,9,10,32,34,51). On the other hand, protein LA0195 failed to bind to the putative promoter region of transcript III, and this fact echoed the weak transcription initiation activity of this region tested in E. coli.…”

Section: Discussionmentioning

confidence: 97%

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Identification of a Novel Prophage-Like Gene Cluster Actively Expressed in Both Virulent and Avirulent Strains of Leptospira interrogans Serovar Lai

Qin

Zhang

et al. 2008

Infect Immun

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DNA microarray analysis was used to compare the differential gene expression profiles between Leptospira interrogans serovar Lai type strain 56601 and its corresponding attenuated strain IPAV. A 22-kb genomic island covering a cluster of 34 genes (i.e., genes LA0186 to LA0219) was actively expressed in both strains but concomitantly upregulated in strain 56601 in contrast to that of IPAV. Reverse transcription-PCR assays proved that the gene cluster comprised five transcripts. Gene annotation of this cluster revealed characteristics of a putative prophage-like remnant with at least 8 of 34 sequences encoding prophage-like proteins, of which the LA0195 protein is probably a putative prophage CI-like regulator. The transcription initiation activities of putative promoter-regulatory sequences of transcripts I, II, and III, all proximal to the LA0195 gene, were further analyzed in the Escherichia coli promoter probe vector pKK232-8 by assaying the reporter chloramphenicol acetyltransferase (CAT) activities. The strong promoter activities of both transcripts I and II indicated by the E. coli CAT assay were well correlated with the in vitro sequence-specific binding of the recombinant LA0195 protein to the corresponding promoter probes detected by the electrophoresis mobility shift assay. On the other hand, the promoter activity of transcript III was very low in E. coli and failed to show active binding to the LA0195 protein in vitro. These results suggested that the LA0195 protein is likely involved in the transcription of transcripts I and II. However, the identical complete DNA sequences of this prophage remnant from these two strains strongly suggests that possible regulatory factors or signal transduction systems residing outside of this region within the genome may be responsible for the differential expression profiling in these two strains. L. weilii, L. fainei, L. inadai, and L. meyeri (1, 24). It affects a wide range of mammalian hosts, and human beings and animals become infected through contact with urine-contaminated soil and water. Because of the large spectra of animal species that serve as reservoirs, leptospirosis is considered to be the most widely spread zoonosis. Leptospirosis is a globally important zoonosis caused by infection of pathogenicProphages are commonly found in sequenced bacterial genomes, many of which appeared to be defective and in a state of mutational decay (15,(18)(19)(20). However, both intact and defective prophages may play a special physiological role in the host bacteria and, thus far, they have been implicated in serotype conversion, pathogenesis, and phage immunity (13,15,16,18,20,22,29,38,53). Genomic sequences of L. interrogans have been determined for two strains of different serotypes (40, 43), but no intact phage sequence was identified in any one of them.In the present study, we compared the gene expression profiles between the virulent L. interrogans serogroup Icterohaemorrhagiae serovar Lai type strain 56601 and its corresponding high-passage avirulent strain IPAV ...

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 97%

Identification of a Novel Prophage-Like Gene Cluster Actively Expressed in Both Virulent and Avirulent Strains of Leptospira interrogans Serovar Lai

Qin

Zhang

et al. 2008

Infect Immun

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“…Orf286 binds speci®cally to a region upstream of its own gene It is known that the genetic switch of several temperate E. coli and LAB phages is transcriptionally controlled by the binding of a phage repressor to speci®c DNA sites upstream of and proximal to its own structural gene (Ptashne 1992;Nauta et al 1996;Ladero et al 1998;Venema et al 1999;Kakikawa et al 2000). For the lactococcal phages, this has only been shown for the r1t and Tuc2009 phage repressors Leach et al 1999).…”

Section: Resultsmentioning

confidence: 99%

“…plantarum (Kodaira et al 1997) and for r1t, BK5-T, Tuc2009, TP901-1, and TPW22 of Lactococcus lactis (van de Guchte et al 1994;Boyce et al 1995b;Nauta et al 1996;Madsen and Hammer 1998;Petersen 1998;Madsen et al 1999). However, only the repressors of the lactococcal phages r1t, Tuc2009, and Tp901-1 (van de Guchte et al 1994;Nauta et al 1996;Madsen et al 1999), and of the lactobacillus phages A2 (Ladero et al 1998;GarcõÂ a et al 1999) and /gle (Kakikawa et al 2000) have been identi®ed and partly characterized.…”

Section: Introductionmentioning

confidence: 99%

Analysis of a regulator involved in the genetic switch between lysis and lysogeny of the temperate Lactococcus lactis phage φLC3

2001

Mol Gen Genomics

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Sequencing of a 1.3-kb fragment of DNA from the temperate Lactococceus lactis subsp. cremoris phage phiLC3 revealed a pair of two divergently oriented ORFs, orf63 and orf286. The deduced amino acid sequence of the product of orf286 showed extensive homology to those of repressors of the temperate lactococcal phages rlt, Tuc2009 and BK5-T. A mutant with an amber mutation in orf286 gave rise to a clear plaque phenotype, indicating that this gene is involved in the lytic and lysogenic development of phiLC3. Gel mobility shift assays showed that the partially purified Orf286 protein bound specifically to the 224-bp intergenic region located between orf286 and orf63, and further characterization by DNase I footprinting analysis revealed that Orf286 protects two distinct sites within this region. Sequence analysis of the intergenic region revealed two putative, divergently oriented promoters, P1 and P2; orf286 and orf63 are probably transcribed from P1 and P2, respectively. In vivo analyses of P1 and P2 using beta-galactosidase as a reporter enzyme in L. lactis showed that transcription from P1 was repressed while transcription from P2 was stimulated in the presence of the Orf286 protein. These results suggest a complex role for the Orf286 protein in regulating the genetic switch between lytic and lysogenic growth of phiLC3.

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“…Both types have been described for phages of either gram-positive or gramnegative bacteria. Phages A2, TP901-1, and others of lactic acid bacteria (2,4,14,17,23,27), L5 of Mycobacterium spp. (3), PBSX of Bacillus subtilis (28), and a number of coliphage can be classified as the "one-immunity-control-region" type.…”

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confidence: 99%

immX Immunity Region of Rhizobium Phage 16 - 3 : Two Overlapping Cistrons of Repressor Function

et al. 2003

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16-3 is a temperate phage of the symbiotic nitrogen-fixing bacterium Rhizobium meliloti 41. Its prophage state and immunity against superinfection by homoimmune phages are governed by a complex set of controls: the immC and immX repressor systems and the avirT element are all located in well-separated, distinct regions which span 25 kb on the bacteriophage chromosome. The anatomy and function of the immC region are well documented; however, fewer analyses have addressed the immX and avirT regions. We focused in this paper on the immX region and dissected it into two major parts: X U/L and X V . The X U/L part (0.6 kb) contained two overlapping cistrons, X U and X L , coding for proteins pXU and pXL, respectively. Inactivation of either gene inactivated the repressor function of the immX region. Loss-of-function mutants of X U and X L complemented each other in trans in double lysogens. The X V part (1 kb) contained a target for X U/L repressor action. Mutations at three sites in X V led to various degree of ImmX insensitivity in a hierarchic manner. Two sites (X V1 and X V3 ) exhibited the inverted-repeat structures characteristic of many repressor binding sites. However, X V1 could also be folded into a transcription terminator. Of the two immunity regions of 16-3, immX seems to be unique both in its complex genetic anatomy and in its sequence. To date, no DNA or peptide sequence homologous to that of ImmX has been found in the data banks. In contrast, immC shares properties of a number of immunity systems commonly found in temperate phages.A bacterium carrying a prophage is immune to superinfection by a homoimmune phage (20). Superinfection immunity is due to the binding of the prophage-encoded repressor protein(s) to specific operator sites of the phage chromosomes. The repressor prevents vegetative phage development by acting on both the resident prophage and the newly injected DNA of a homoimmune phage (34). Although the repressor systems may differ in specificity, topology, and position of the relevant genes on the genetic map, two major types are emerging among temperate phages investigated by genetic analyses: (i) phages with one immunity control region (canonized by coliphage ) (20, 34) and (ii) phages with dual control regions (the archetype is Salmonella phage P22) (44). Both types have been described for phages of either gram-positive or gramnegative bacteria. Phages A2, TP901-1, and others of lactic acid bacteria (2,4,14,17,23,27), L5 of Mycobacterium spp. (3), PBSX of Bacillus subtilis (28), and a number of coliphage can be classified as the "one-immunity-control-region" type. In contrast, Escherichia coli phage 186 and 105 of B. subtilis possess two immunity regions, like P22 (9, 10, 22, 41). The highly complex P22 immI region encodes an antagonist (Ant) of the C2 immunity repressor and proteins and RNAs that control ant expression (mnt, arc, and sar) (37). An even more complex immunity system is represented by phage P1, where three immunity regions function in a complex network (19).The superinfe...

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The genetic switch for the regulatory pathway of Lactobacillus plantarum phage φg1e: characterization of the promoter PL, the repressor gene cpg, and the cpg-encoded protein Cpg in Escherichia coli

Cited by 18 publications

References 27 publications

Identification of a Novel Prophage-Like Gene Cluster Actively Expressed in Both Virulent and Avirulent Strains of Leptospira interrogans Serovar Lai

Identification of a Novel Prophage-Like Gene Cluster Actively Expressed in Both Virulent and Avirulent Strains of Leptospira interrogans Serovar Lai

Analysis of a regulator involved in the genetic switch between lysis and lysogeny of the temperate Lactococcus lactis phage φLC3

immX Immunity Region of Rhizobium Phage 16 - 3 : Two Overlapping Cistrons of Repressor Function

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