Genetic characterization of Pseudomonas aeruginosa bacteriophage KPP10

Uchiyama, Jumpei; Rashel, Mohammad; Takemura, Iyo; Kato, Shin‐ichiro; Ujihara, Takako; Muraoka, Asako; Matsuzaki, Shigenobu; Daibata, Masanori

doi:10.1007/s00705-011-1210-x

Cited by 20 publications

(19 citation statements)

References 27 publications

(25 reference statements)

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“…A virulent Pseudomonas phage belonging to the family Siphoviridae is proposed as a therapeutic phage (5). Phage KPP23 was isolated from a water sample collected from Urado Bay, Kochi City, Kochi, Japan, using P. aeruginosa strain PA101 as the host strain (6). According to the morphological analysis by transmission electron microscopy, phage KPP23 has an icosahedral head and a flexible tail (mean ± standard deviation, 67.3 ± 4.5 nm in diameter and 141.4 ± 11.8 nm in length, respectively; n = 20), and was classified under the family Siphoviridae .…”

Section: Genome Announcementmentioning

confidence: 99%

Genome Analysis of Pseudomonas aeruginosa Bacteriophage KPP23, Belonging to the Family Siphoviridae

et al. 2014

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Section: Genome Announcementmentioning

confidence: 99%

Genome Analysis of Pseudomonas aeruginosa Bacteriophage KPP23, Belonging to the Family Siphoviridae

et al. 2014

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“…Genomic DNA was extracted from the phage particles essentially as described elsewhere (43,44). Moreover, H. pylori strain NY43, which releases phage KHP30, was cultured in BEV medium and then pelleted.…”

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confidence: 99%

“…Phage proteins were prepared from the phage purified by CsCl density gradient ultracentrifugation for SDS-PAGE, as described previously (44). The phage proteins were separated electrophoretically on a 12.5% SDS-polyacrylamide gel with an (unstained) XLLadder Broad marker (APRO Life Science Institute, Tokushima, Japan), basically followed by the protocol described elsewhere (49).…”

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confidence: 99%

Characterization of Helicobacter pylori Bacteriophage KHP30

Uchiyama

Takeuchi

Kato

et al. 2013

Appl Environ Microbiol

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Helicobacter pylori inhabits the stomach mucosa and is a causative agent of stomach ulcer and cancer. In general, bacteriophages (phages) are strongly associated with bacterial evolution, including the development of pathogenicity. Several tailed phages have so far been reported in H. pylori. We have isolated an H. pylori phage, KHP30, and reported its genomic sequence. In this study, we examined the biological characteristics of phage KHP30. Phage KHP30 was found to be a spherical lipid-containing phage with a diameter of ca. 69 nm. Interestingly, it was stable from pH 2.5 to pH 10, suggesting that it is adapted to the highly acidic environment of the human stomach. Phage KHP30 multiplied on 63.6% of clinical H. pylori isolates. The latent period was ca. 140 min, shorter than the doubling time of H. pylori (ca. 180 min). The burst size was ca. 13, which was smaller than the burst sizes of other known tailed or spherical phages. Phage KHP30 seemed to be maintained as an episome in H. pylori strain NY43 cells, despite a predicted integrase gene in the KHP30 genomic sequence. Seven possible virion proteins of phage KHP30 were analyzed using N-terminal protein sequencing and mass spectrometry, and their genes were found to be located on its genomic DNA. The genomic organization of phage KHP30 differed from the genomic organizations in the known spherical phage families Corticoviridae and Tectiviridae. This evidence suggests that phage KHP30 is a new type of spherical phage that cannot be classified in any existing virus category. Helicobacter pylori, a Gram-negative spiral bacterium, colonizes the human stomach mucosa (1). It causes chronic inflammation, which may progress to peptic ulceration, atrophic gastritis, and gastric cancer (2). Recent studies of H. pylori have revealed that the pathogenicity of H. pylori strains is related to the specific geographic region from which they derive (3, 4). An East Asian type of H. pylori strain most likely causes gastric cancer and highly likely contains cytotoxin-associated gene A (cagA), a carcinogenic genetic element (1, 3). However, how the pathogenicity of the H. pylori strains has evolved is unknown, although H. pylori inhabits a very restricted niche in the stomach mucosa. Therefore, the factors involved in H. pylori evolution must be investigated.Bacteriophages (phages) are the most diverse and abundant life form on Earth. Phages practice lateral gene transfer and are involved in a coevolutionary arms race with bacteria (5-8), so they may well be factors involved in bacterial evolution. Several H. pylori phages have been reported (9-13), and some of these have been morphologically studied and have been found to belong to the order Caudovirales (i.e., the tailed phages) (9-12). Recent advances in sequencing technologies have allowed intensive genomic analyses of Helicobacter spp. and have also suggested the presence of phages in the bacterial genome (10,11,14,15). However, the biological characteristics of these H. pylori phages are not well-known.We have recently iso...

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“…The lytic activity (i.e., lysis from within, lysis from without, or no lysis) of phage KPP22 against clinically isolated P. aeruginosa strains that included various serotypes was compared with that of phage KPP12 (16,21,31). Phage KPP22 showed lysis from within on 92.1% (35 of 38) of the tested clinically isolated strains, while phage KPP12 showed lysis from within and lysis from without on 68.4% (26 of 38) of the tested clinically isolated strains (Table 1).…”

Section: Resultsmentioning

confidence: 99%

“…The P. aeruginosa strains used in this study, which had been isolated from the patients in Kochi, Japan, and have been described previously, are listed in Table 1 (14,21). Phage KPP22 was isolated using strain PA33 as the host by a method described elsewhere (22).…”

Section: Methodsmentioning

confidence: 99%

Analyses of Short-Term Antagonistic Evolution of Pseudomonas aeruginosa Strain PAO1 and Phage KPP22 (Myoviridae Family, PB1-Like Virus Genus)

Uchiyama

Suzuki

Nishifuji

et al. 2016

Appl Environ Microbiol

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Pseudomonas aeruginosa causes serious intractable infections in humans and animals. Bacteriophage (phage) therapy has been applied to treat P. aeruginosa infections, and phages belonging to the PB1-like virus genus in the Myoviridae family have been used as therapeutic phages. To achieve safer and more effective phage therapy, the use of preadapted phages is proposed. To understand in detail such phage preadaptation, the short-term antagonistic evolution of bacteria and phages should be studied. In this study, the short-term antagonistic evolution of bacteria and PB1-like phage was examined by studying phage-resistant clones of P. aeruginosa strain PAO1 and mutant PB1-like phages that had recovered their infectivity. First, phage KPP22 was isolated and characterized; it was classified as belonging to the PB1-like virus genus in the Myoviridae family. Subsequently, three KPP22-resistant PAO1 clones and three KPP22 mutant phages capable of infecting these clones were isolated in three sets of in vitro experiments. It was shown that the bacterial resistance to phage KPP22 was caused by significant decreases in phage adsorption and that the improved infectivity of KPP22 mutant phages was caused by significant increases in phage adsorption. The KPP22-resistant PAO1 clones and the KPP22 mutant phages were then analyzed genetically. All three KPP22-resistant PAO1 clones, which were deficient for the O5 antigen, had a common nonsense mutation in the wzy gene. All the KPP22 mutant phage genomes showed the same four missense mutations in the open reading frames orf060, orf065, and orf086. The information obtained in this study should be useful for further development of safe and efficient phage therapy. IMPORTANCEPseudomonas aeruginosa causes serious intractable infections in humans and animals; bacteriophage (phage) therapy has been utilized to treat P. aeruginosa infections, and phages that belong to the PB1-like virus genus in the family Myoviridae have been used as therapeutic phages. The preadapted phage is trained in advance through the antagonistic evolution of bacteria and phage and is proposed to be used to achieve safer and more effective phage therapy. In this study, to understand the phage preadaptation, the in vitro short-term antagonistic evolution was studied using P. aeruginosa strain PAO1 and the newly isolated PB1-like phage KPP22. Phage KPP22 was characterized, and the molecular framework regarding the phage preadaptation of KPP22 was elucidated. The importance of study of antagonistic evolution of bacteria and phage in phage therapy is discussed.

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Genetic characterization of Pseudomonas aeruginosa bacteriophage KPP10

Cited by 20 publications

References 27 publications

Genome Analysis of Pseudomonas aeruginosa Bacteriophage KPP23, Belonging to the Family Siphoviridae

Genome Analysis of Pseudomonas aeruginosa Bacteriophage KPP23, Belonging to the Family Siphoviridae

Characterization of Helicobacter pylori Bacteriophage KHP30

Analyses of Short-Term Antagonistic Evolution of Pseudomonas aeruginosa Strain PAO1 and Phage KPP22 (Myoviridae Family, PB1-Like Virus Genus)

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