The
 repA
 Gene of the Linear
 Yersinia enterocolitica
 Prophage PY54 Functions as a Circular Minimal Replicon in
 Escherichia coli

Ziegelin, Günter; Tegtmeyer, Nicole; Hertwig, Stefan; Hammerl, Jens A.; Appel, Bernd; Lanka, Erich

doi:10.1128/jb.187.10.3445-3454.2005

Cited by 14 publications

(10 citation statements)

References 41 publications

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“…Similar to lambda, binding was observed to three operators ( Figure 2 B) situated between cB and cro . In addition, two operators (O L ) overlapping with the predicted promoter of the plasmid replication gene repA were bound [ 9 ]. N15 CB was therefore suggested to be implicated in both the regulatory circuitry of phage propagation and the control of plasmid replication [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

The Molecular Switch of Telomere Phages: High Binding Specificity of the PY54 Cro Lytic Repressor to a Single Operator Site

Hammerl

Roschanski²,

Johne

et al. 2015

Viruses

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Temperate bacteriophages possess a molecular switch, which regulates the lytic and lysogenic growth. The genomes of the temperate telomere phages N15, PY54 and ϕKO2 harbor a primary immunity region (immB) comprising genes for the prophage repressor, the lytic repressor and a putative antiterminator. The roles of these products are thought to be similar to those of the lambda proteins CI, Cro and Q, respectively. Moreover, the gene order and the location of several operator sites in the prototype telomere phage N15 and in ϕKO2 are also reminiscent of lambda-like phages. By contrast, in silico analyses revealed the presence of only one operator (OR3) in PY54. The purified PY54 Cro protein was used for EMSA studies demonstrating that it exclusively binds to a 16-bp palindromic site (OR3) upstream of the prophage repressor gene. The OR3 operator sequences of PY54 and ϕKO2/N15 only differ by their peripheral base pairs, which are responsible for Cro specificity. PY54 cI and cro transcription is regulated by highly active promoters initiating the synthesis of a homogenious species of leaderless mRNA. The location of the PY54 Cro binding site and of the identified promoters suggests that the lytic repressor suppresses cI transcription but not its own synthesis. The results indicate an unexpected diversity of the growth regulation mechanisms in lambda-related phages.

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Section: Introductionmentioning

confidence: 99%

The Molecular Switch of Telomere Phages: High Binding Specificity of the PY54 Cro Lytic Repressor to a Single Operator Site

Hammerl

Roschanski²,

Johne

et al. 2015

Viruses

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“…The arrangement of genes for structural proteins, partitioning proteins, the protelomerase including its target site tel, the replication protein repA and lysis proteins is so similar that it appears likely that these phages share a common ancestor. 3,20 At first glance this also applies to the primary immunity region immB of the telomere phages (Fig. 2), which resembles the genetic switch of many lambdoid phages.…”

Section: Discussionmentioning

confidence: 92%

The diverse genetic switch of enterobacterial and marine telomere phages

et al. 2016

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Temperate bacteriophages possess a genetic switch which regulates the lytic and lysogenic cycle. The genomes of the enterobacterial telomere phages N15, PY54 and fKO2 harbor a primary immunity region (immB) comprising genes for the prophage repressor, the lytic repressor and a putative antiterminator, similar to CI, Cro and Q of lambda, respectively. Moreover, N15 and fKO2 contain 3 related operator (OR) sites between cI and cro, while only one site (O R 3) has been detected in PY54. Marine telomere phages possess a putative cI gene but not a cro-like gene. Instead, a gene is located at the position of cro, whose product shows some similarity to the PY54 ORF42 product, the function of which is unknown. We have determined the transcription start sites of the predicted repressor genes of N15, PY54, fKO2 and of the marine telomere phage VP58.5. The influence of the genes on phage propagation was analyzed in E. coli, Y. enterocolitica and V. parahaemolyticus. We show that the repressors and antiterminators of N15, fKO2 and PY54 exerted their predicted activities. However, while the proteins of both N15 and fKO2 affected lysis and lysogeny by N15, they did not affect PY54 propagation. On the other hand, the respective PY54 proteins exclusively influenced the propagation of this phage. The immB region of VP58.5 contains 2 genes that revealed prophage repressor activity, while a lytic repressor gene could not be identified. The results indicate an unexpected diversity of the growth regulation mechanisms in these temperate phages.

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“…The predicted RepA proteins of the phages are 96% identical. The construction of the replicative circular miniplasmid pJH1201 comprising a 4.8-kb restriction fragment of the Vp58.5 prophage indicated that the origin of this phage is located close to or even within repA, similarly to the replication genes of N15, PY54, and ⌽KO2 (35,42). Plasmid pJH1201 coexists with the aforementioned prophages in the same cell.…”

Section: Discussionmentioning

confidence: 99%

“…Besides tel, all telomere phages possess the gene repA, encoding a multifunctional replication protein. repA of N15 and PY54 was shown to harbor the prophage replication origin and to function as a circular minimal replicon (35,42). Compatibility studies demonstrated that the N15 and ⌽KO2 plasmids belong to the same incompatibility group, whereas the PY54 plasmid is able to coexist with these two prophages in doubly lysogenic E. coli and Y. enterocolitica hosts (19).…”

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confidence: 99%

The Linear Plasmid Prophage Vp58.5 of Vibrio parahaemolyticus Is Closely Related to the Integrating Phage VHML and Constitutes a New Incompatibility Group of Telomere Phages

Zabala

Hammerl

Espejo

et al. 2009

J Virol

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Vibrio parahaemolyticus O3:K6 pandemic strains recovered in Chile frequently possess a 42-kb plasmid which is the prophage of a myovirus. We studied the prototype phage VP58.5 and show that it does not integrate into the host cell chromosome but replicates as a linear plasmid (Vp58.5) with covalently closed ends (telomeres). The Vp58.5 replicon coexists with other plasmid prophages (N15, PY54, and ⌽KO2) in the same cell and thus belongs to a new incompatibility group of telomere phages. We determined the complete nucleotide sequence (42,612 nucleotides) of the VP58.5 phage DNA and compared it with that of the plasmid prophage. The two molecules share the same nucleotide sequence but are 35% circularly permuted to each other. In contrast to the hairpin ends of the plasmid, VP58.5 phage DNA contains 5-protruding ends. The VP58.5 sequence is 92% identical to the sequence of phage VHML, which was reported to integrate into the host chromosome. However, the gene order and termini of the phage DNAs are different. The VHML genome exhibits the same gene order as does the Vp58.5 plasmid. VHML phage DNA has been reported to contain terminal inverted repeats. This repetitive sequence is similar to the telomere resolution site (telRL) of VP58.5 which, after processing by the phage protelomerase, forms the hairpin ends of the Vp58.5 prophage. It is discussed why these closely related phages may be so different in terms of their genome ends and their lifestyle.Most temperate bacteriophages integrate into the host chromosome during lysogeny. However, there are some phages (telomere phages) whose prophages are linear plasmids with covalently closed ends. Members of this group of phages are the siphoviruses N15, PY54, and ⌽KO2 isolated from Escherichia coli, Yersinia enterocolitica, and Klebsiella oxytoca, respectively, and the recently described myoviruses ⌽HAP-1 of Halomonas aquamarina and VP882 of Vibrio parahaemolyticus (6,20,23,26,37). Despite their different origins (enterobacteria versus marine bacterium) and morphologies, all known telomere phages share similar genome organizations and some protein similarities. The linear DNA of each phage is a circular permutation of the respective linear plasmid prophage. For the generation of the terminal hairpins of the linear plasmid, the protelomerase (Tel) is essential (8). This enzyme has cleaving/joining activity; its target is a large palindromic DNA sequence called the telomere resolution site (telRL) located upstream of tel on the phage genome. After cleaving telRL by staggered cuts, the resulting self-complementary single-stranded DNA overhangs fold back and are rejoined by the protelomerase (9). Besides tel, all telomere phages possess the gene repA, encoding a multifunctional replication protein. repA of N15 and PY54 was shown to harbor the prophage replication origin and to function as a circular minimal replicon (35,42). Compatibility studies demonstrated that the N15 and ⌽KO2 plasmids belong to the same incompatibility group, whereas the PY54 plasmid is able to coexist...

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The repA Gene of the Linear Yersinia enterocolitica Prophage PY54 Functions as a Circular Minimal Replicon in Escherichia coli

Cited by 14 publications

References 41 publications

The Molecular Switch of Telomere Phages: High Binding Specificity of the PY54 Cro Lytic Repressor to a Single Operator Site

The Molecular Switch of Telomere Phages: High Binding Specificity of the PY54 Cro Lytic Repressor to a Single Operator Site

The diverse genetic switch of enterobacterial and marine telomere phages

The Linear Plasmid Prophage Vp58.5 of Vibrio parahaemolyticus Is Closely Related to the Integrating Phage VHML and Constitutes a New Incompatibility Group of Telomere Phages

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