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1.
Viruses ; 13(2)2021 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-33498475

RESUMO

The paper covers the history of the discovery and description of phiKZ, the first known giant bacteriophage active on Pseudomonas aeruginosa. It also describes its unique features, especially the characteristic manner of DNA packing in the head around a cylinder-shaped structure ("inner body"), which probably governs an ordered and tight packaging of the phage genome. Important properties of phiKZ-like phages include a wide range of lytic activity and the blue opalescence of their negative colonies, and provide a background for the search and discovery of new P. aeruginosa giant phages. The importance of the phiKZ species and of other giant phage species in practical phage therapy is noted given their broad use in commercial phage preparations.


Assuntos
Genoma Viral , Fagos de Pseudomonas/genética , Fagos de Pseudomonas/fisiologia , Pseudomonas aeruginosa/virologia , Microscopia Crioeletrônica , Terapia por Fagos , Filogenia , Fagos de Pseudomonas/ultraestrutura
2.
Sci Rep ; 7(1): 13731, 2017 10 23.
Artigo em Inglês | MEDLINE | ID: mdl-29062062

RESUMO

The Red Queen hypothesis posits that antagonistic co-evolution between interacting species results in recurrent natural selection via constant cycles of adaptation and counter-adaptation. Interactions such as these are at their most profound in host-parasite systems, with bacteria and their viruses providing the most intense of battlefields. Studies of bacteriophage evolution thus provide unparalleled insight into the remarkable elasticity of living entities. Here, we report a novel phenomenon underpinning the evolutionary trajectory of a group of dsDNA bacteriophages known as the phiKMVviruses. Employing deep next generation sequencing (NGS) analysis of nucleotide polymorphisms we discovered that this group of viruses generates enhanced intraspecies heterogeneity in their genomes. Our results show the localisation of variants to genes implicated in adsorption processes, as well as variation of the frequency and distribution of SNPs within and between members of the phiKMVviruses. We link error-prone DNA polymerase activity to the generation of variants. Critically, we show trans-activity of this phenomenon (the ability of a phiKMVvirus to dramatically increase genetic variability of a co-infecting phage), highlighting the potential of phages exhibiting such capabilities to influence the evolutionary path of other viruses on a global scale.


Assuntos
Bacteriófagos/genética , DNA Viral/genética , Evolução Molecular , Bacteriófagos/enzimologia , DNA Polimerase Dirigida por DNA/química , DNA Polimerase Dirigida por DNA/metabolismo , Sequenciamento de Nucleotídeos em Larga Escala , Modelos Moleculares , Fenótipo , Conformação Proteica , Análise de Sequência de DNA
3.
Front Microbiol ; 7: 1631, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27790211

RESUMO

This review discusses the potential application of bacterial viruses (phage therapy) toward the eradication of antibiotic resistant Pseudomonas aeruginosa in children with cystic fibrosis (CF). In this regard, several potential relationships between bacteria and their bacteriophages are considered. The most important aspect that must be addressed with respect to phage therapy of bacterial infections in the lungs of CF patients is in ensuring the continuity of treatment in light of the continual occurrence of resistant bacteria. This depends on the ability to rapidly select phages exhibiting an enhanced spectrum of lytic activity among several well-studied phage groups of proven safety. We propose a modular based approach, utilizing both mono-species and hetero-species phage mixtures. With an approach involving the visual recognition of characteristics exhibited by phages of well-studied phage groups on lawns of the standard P. aeruginosa PAO1 strain, the simple and rapid enhancement of the lytic spectrum of cocktails is permitted, allowing the development of tailored preparations for patients capable of circumventing problems associated with phage resistant bacterial mutants.

4.
Genome Announc ; 4(2)2016 Mar 03.
Artigo em Inglês | MEDLINE | ID: mdl-26941153

RESUMO

The complete genome of the Pseudomonas aeruginosa bacteriophage PM105 is 39,593 bp long. The phage belongs to the B3 family of transposable Mu-like phages, as confirmed by the presence of bacterial DNA joined to the phage genome ends. PM105, together with other B3-like phages, form a newly arising species.

5.
FEMS Microbiol Lett ; 362(9)2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25825475

RESUMO

A complete nucleotide sequence of the new Pseudomonas aeruginosa Luz24likevirus phiCHU was obtained. This virus was shown to have a unique host range whereby it grew poorly on the standard laboratory strain PAO1, but infected 26 of 46 clinical isolates screened, and strains harbouring IncP2 plasmid pMG53. It was demonstrated that phiCHU has single-strand interruptions in its genome. Analysis of the phiCHU genome also suggested that recombination event(s) participated in the evolution of the leftmost portion of the genome, presumably encoding early genes.


Assuntos
Genoma Viral , Especificidade de Hospedeiro , Podoviridae/genética , Fagos de Pseudomonas/genética , Pseudomonas aeruginosa/virologia , Sequência de Bases , Dados de Sequência Molecular , Fases de Leitura Aberta , Plasmídeos , Análise de Sequência de DNA
6.
Virol Sin ; 30(1): 33-44, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25680443

RESUMO

The emergence of multidrug-resistant bacterial pathogens forced us to consider the phage therapy as one of the possible alternative approaches to treatment. The purpose of this paper is to consider the conditions for the safe, long-term use of phage therapy against various infections caused by Pseudomonas aeruginosa. We describe the selection of the most suitable phages, their most effective combinations and some approaches for the rapid recognition of phages unsuitable for use in therapy. The benefits and disadvantages of the various different approaches to the preparation of phage mixtures are considered, together with the specific conditions that are required for the safe application of phage therapy in general hospitals and the possibilities for the development of personalized phage therapy.


Assuntos
Bacteriófagos/fisiologia , Terapia Biológica , Infecções por Pseudomonas/terapia , Pseudomonas aeruginosa/virologia , Bacteriófagos/classificação , Bacteriófagos/genética , Humanos , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/fisiologia
7.
Viruses ; 5(1): 15-53, 2012 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-23344559

RESUMO

Pseudomonas aeruginosa is a frequent participant in wound infections. Emergence of multiple antibiotic resistant strains has created significant problems in the treatment of infected wounds. Phage therapy (PT) has been proposed as a possible alternative approach. Infected wounds are the perfect place for PT applications, since the basic condition for PT is ensured; namely, the direct contact of bacteria and their viruses. Plenty of virulent ("lytic") and temperate ("lysogenic") bacteriophages are known in P. aeruginosa. However, the number of virulent phage species acceptable for PT and their mutability are limited. Besides, there are different deviations in the behavior of virulent (and temperate) phages from their expected canonical models of development. We consider some examples of non-canonical phage-bacterium interactions and the possibility of their use in PT. In addition, some optimal approaches to the development of phage therapy will be discussed from the point of view of a biologist, considering the danger of phage-assisted horizontal gene transfer (HGT), and from the point of view of a surgeon who has accepted the Hippocrates Oath to cure patients by all possible means. It is also time now to discuss the possible approaches in international cooperation for the development of PT. We think it would be advantageous to make phage therapy a kind of personalized medicine.


Assuntos
Terapia Biológica/métodos , Infecções por Pseudomonas/terapia , Fagos de Pseudomonas/genética , Pseudomonas aeruginosa/virologia , Infecção dos Ferimentos/terapia , Humanos , Biologia Molecular/métodos , Fagos de Pseudomonas/crescimento & desenvolvimento
8.
J Mol Biol ; 354(3): 536-45, 2005 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-16256135

RESUMO

Pseudomonas aeruginosa phage EL is a dsDNA phage related to the giant phiKZ-like Myoviridae. The EL genome sequence comprises 211,215 bp and has 201 predicted open reading frames (ORFs). The EL genome does not share DNA sequence homology with other viruses and micro-organisms sequenced to date. However, one-third of the predicted EL gene products (gps) shares similarity (Blast alignments of 17-55% amino acid identity) with phiKZ proteins. Comparative EL and phiKZ genomics reveals that these giant phages are an example of substantially diverged genetic mosaics. Based on the position of similar EL and phiKZ predicted gene products, five genome regions can be delineated in EL, four of which are relatively conserved between EL and phiKZ. Region IV, a 17.7 kb genome region with 28 predicted ORFs, is unique to EL. Fourteen EL ORFs have been assigned a putative function based on protein similarity. Assigned proteins are involved in DNA replication and nucleotide metabolism (NAD+-dependent DNA ligase, ribonuclease HI, helicase, thymidylate kinase), host lysis and particle structure. EL-gp146 is the first chaperonin GroEL sequence identified in a viral genome. Besides a putative transposase, EL harbours predicted mobile endonucleases related to H-N-H and LAGLIDADG homing endonucleases associated with group I intron and intein intervening sequences.


Assuntos
Genoma Viral/genética , Fagos de Pseudomonas/genética , Pseudomonas aeruginosa/virologia , Sequência de Aminoácidos , Replicação do DNA/genética , Endonucleases/metabolismo , Genes Virais/genética , Inteínas , Microscopia Eletrônica , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Fagos de Pseudomonas/ultraestrutura , Pseudomonas aeruginosa/ultraestrutura , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Transcrição Gênica/genética , Proteínas Virais/genética , Ativação Viral
9.
Res Microbiol ; 154(4): 269-75, 2003 May.
Artigo em Inglês | MEDLINE | ID: mdl-12798231

RESUMO

Recently we have accomplished the entire DNA sequence of bacteriophage phiKZ, a giant virus infecting Pseudomonas aeruginosa. The 280334-bp of phiKZ genome is a linear, circularly permutated and terminally redundant, AT-rich dsDNA molecule that contains no sites for NotI, PstI, SacI, SmaI, XhoI and XmaIII endonucleases. Limited homology to other bacteriophages on the DNA and protein levels indicated that phiKZ represents a distinct branch of the Myoviridae family. In this work, we analyzed a group of six P. aeruginosa phages (Lin68, Lin21, PTB80, NN, EL, and RU), which are morphologically similar to phiKZ, have similar genome size and low G+C content. All phages have a broad host range among P. aeruginosa strains, and they are resistant to the inhibitory action of many P. aeruginosa plasmids. The analysis of the genomic DNA by restriction enzymes and DNA-DNA hybridization shows that phages are representative of three phiKZ-like species: phiKZ-type (phiKZ, Lin21, NN and PTB80), EL-type (EL and RU) and Lin68 which has a shorter tail than other phages. Except for related phages EL and RU, all phiKZ-like phages have identical N-terminal amino acid sequences of the major capsid protein. Random genome sequencing shows that the EL and RU phages have no homology to the phiKZ-like phages on DNA level. We propose that the phiKZ, Lin21, NN, PTB80 and Lin68 phages can be included in a new phiKZ genus, and that the EL and RU phages belong to a separate genus within the Myoviridae family. Based on the resistance to many restriction enzymes and the transduction ability, there are indications that over the long pathway of evolution, the phiKZ-like phages probably inherited the capacity to infect different bacterial species.


Assuntos
Evolução Biológica , Myoviridae/classificação , Fagos de Pseudomonas/classificação , Pseudomonas aeruginosa/virologia , DNA Viral/análise , DNA Viral/isolamento & purificação , Genoma Viral , Myoviridae/química , Myoviridae/genética , Myoviridae/ultraestrutura , Fagos de Pseudomonas/química , Fagos de Pseudomonas/genética , Fagos de Pseudomonas/ultraestrutura , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico , Proteínas Virais/análise
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