Bacteriophage resistance inLactococcus

Dinsmore, Polly K.; Klaenhammer, Todd R.

doi:10.1007/bf02779022

Cited by 57 publications

(26 citation statements)

References 106 publications

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“…Bacteriophages can contain heavily modified nucleic acid in order to evade host restriction and modification systems (Bickle & Krüger, 1993;Dinsmore & Klaenhammer, 1995). QPA3 DNA was isolated and proved readily digestable with ApaI, BamHI, BglII, EcoRI, EcoRV and NheI, while EagI, NotI, PmeI, SphI and XbaI were unable to digest QPA3 DNA (data not shown).…”

Section: Qpa3 Genome Sequencingmentioning

confidence: 99%

The Pseudomonas aeruginosa generalized transducing phage φPA3 is a new member of the φKZ-like group of ‘jumbo’ phages, and infects model laboratory strains and clinical isolates from cystic fibrosis patients

et al. 2011

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The Pseudomonas aeruginosa generalized transducing phage QPA3 is a new member of the QKZ-like group of 'jumbo' phages, and infects model laboratory strains and clinical isolates from cystic fibrosis patients Pseudomonas aeruginosa is an important pathogen in cystic fibrosis patients, and a model organism for the study of nosocomially acquired infections, biofilms and intrinsic multidrug resistance. In this study we characterize QPA3, a new generalized transducing bacteriophage for P. aeruginosa. QPA3 transduced chromosomal mutations between PAO1 strains, and infected multiple P. aeruginosa clinical isolates as well as the P. aeruginosa model laboratory strains PAK and PA14. Electron microscopy imaging was used to classify QPA3 in the order Caudovirales and the family Myoviridae. The genome of QPA3 was sequenced and found to contain 309 208 bp, the second-largest bacteriophage currently deposited in GenBank. The genome contains 378 ORFs and five tRNAs. Many ORF products in the QPA3 genome are similar to proteins encoded by P. aeruginosa phage QKZ and Pseudomonas chlororaphis phage 201Q2-1, and so QPA3 was classified genetically as a member of the QKZ-like group of phages. This is the first report of a member of this group of phages acting as a generalized transducer. Given its wide host range, high transduction efficiency and large genome size, the 'jumbo' phage QPA3 could be a powerful tool in functional genomic analysis of diverse P. aeruginosa strains of fundamental and clinical importance. INTRODUCTIONPseudomonas aeruginosa is an opportunistic human pathogen and a leading cause of hospital-acquired infections (Trautmann et al., 2005). The organism primarily infects those with compromised immune systems. Individuals are most susceptible to infection when suffering from open wounds and chronic pulmonary conditions, such as cystic fibrosis (Harrison, 2007;Wagner & Iglewski, 2008). P. aeruginosa has a large genome, with over 5500 genes, and produces a wide variety of secondary metabolites, allowing it to survive in metabolically diverse environments, including soil, water, plants, insects and animals (Boman et al., 1972;Mahajan-Miklos et al., 2000;Stover et al., 2000). The organism can live as biofilms, colonies or as planktonic cells (Tolker-Nielsen & Molin, 2004;Wagner & Iglewski, 2008). Due to its metabolic diversity and its importance as a human pathogen, P. aeruginosa has become an organism of intense study, so genetic tools for this organism have become increasingly important.Generalized transducing bacteriophages are useful genetic tools for strain engineering and for functional genomic analysis in strains that are not naturally competent, such as P. aeruginosa (Wong et al., 2000). Several P. aeruginosa generalized transducing bacteriophages have been described, though the transduction efficiency of these bacteriophages, in particular DMS3, UT1 and F116, can be very low (Budzik et al., 2004;Kidambi et al., 1994;Ripp et al., 1994). Given such low transduction efficiency, occasional background spontaneous mu...

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Section: Qpa3 Genome Sequencingmentioning

confidence: 99%

The Pseudomonas aeruginosa generalized transducing phage φPA3 is a new member of the φKZ-like group of ‘jumbo’ phages, and infects model laboratory strains and clinical isolates from cystic fibrosis patients

et al. 2011

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“…The majority of bacteria carry plasmids or lysogenized phage that protect their host by expressing potent activities that prevent infection by other phages (for reviews, see Dinsmore & Klaenhammer, 1995 ;Snyder, 1995). This is particularly relevant for transduction of environmental P. aeruginosa strains since 40 % of isolates recovered from natural ecosystems (lake water, sediment, soil and sewage) contain DNA sequences homologous to phage genomes (Ogunseitan et al, 1992).…”

Section: The Delivery Vehicle Can Transduce Many Gramnegative Speciesmentioning

confidence: 99%

Development of a P1 phagemid system for the delivery of DNA into Gram-negative bacteria

et al. 2002

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The inability to transform many clinically important Gram-negative bacteria has hampered genetic studies addressing the mechanism of bacterial pathogenesis. This report describes the development and construction of a delivery system utilizing the broad-host-range transducing bacteriophage P1. The phagemids used in this system contain a P1 pac initiation site to package the vector, a P1 lytic replicon to generate concatemeric DNA, a broad-hostrange origin of replication and an antibiotic-resistance determinant to select bacterial clones containing the recircularized phagemid. Phagemid DNA was successfully introduced by infection and stably maintained in members of the families Enterobacteriaceae (Escherichia coli, Shigella flexneri, Shigella dysenteriae, Klebsiella pneumoniae and Citrobacter freundii ) and Pseudomonadaceae (Pseudomonas aeruginosa). In addition to laboratory strains, these virions were used successfully to deliver phagemids to a number of strains isolated from patients. This ability to deliver genetic information to wild-type strains raises the potential for use in antimicrobial therapies and DNA vaccine development.

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“…Bacteriophages continue to be a significant economic problem for the dairy industry (9,30). Starter culture strains which contain naturally occurring phage defense mechanisms can control the problem if they are used in conjunction with sanitation measures.…”

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confidence: 99%

Genetic Analysis of Chromosomal Regions of Lactococcus lactis Acquired by Recombinant Lytic Phages

Durmaz

Klaenhammer

2000

Appl Environ Microbiol

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Recombinant phages are generated when Lactococcus lactis subsp. lactis harboring plasmids encoding the abortive type (Abi) of phage resistance mechanisms is infected with small isometric phages belonging to the P335 species. These phage variants are likely to be an important source of virulent new phages that appear in dairy fermentations. They are distinguished from their progenitors by resistance to Abi defenses and by altered genome organization, including regions of L. lactis chromosomal DNA. The objective of this study was to characterize four recombinant variants that arose from infection of L. lactis NCK203 (Abi ؉ ) with phage 31. HindIII restriction maps of the variants (31.1, 31.2, 31.7, and 31.8) were generated, and these maps revealed the regions containing recombinant DNA. The recombinant region of phage 31.1, the variant that occurred most frequently, was sequenced and revealed 7. Bacteriophages continue to be a significant economic problem for the dairy industry (9, 30). Starter culture strains which contain naturally occurring phage defense mechanisms can control the problem if they are used in conjunction with sanitation measures. Bacteriophages, nevertheless, have the capacity to evolve and rapidly overcome host defense mechanisms (9,13,22,29). The logical routes of phage evolution include point mutations, deletions, and acquisition of new DNA. Coinfecting phages and the genomic contents of the host cell, which can include functional or defective prophages, are potential sources of new DNA. Identification of the genetic routes by which phages adapt and evolve against industrial starter cultures will be an important part of controlling the appearance of new phages in fermentation environments.Lactococcal phages are classified into 12 species based on morphology and DNA homologies (20). The c2 (prolateheaded) species and the 936 and P335 (small isometricheaded) species are the most important taxa, since these are the major organisms that disrupt dairy fermentations worldwide. While the 936 species is composed of only lytic phages, P335 species exhibit high levels of DNA homology between temperate and lytic members (17,20). P335 phages have been appearing in cheese plants with increasing frequency in recent years and are now considered members of an important new phage species (1,17,21,28).Workers in our laboratory have previously characterized a number of phages belonging to the P335 species which are virulent for Lactococcus lactis NCK203. In response to the inhibitory pressure of an abortive type of defense (AbiC) (12), phage ul36 can acquire chromosomal sequences from the host in a recombinational process that generates a related but new virulent phage, ul37 (8,29). Phage ul37 is not inhibited by AbiC and differs in its base plate and tail morphology. The nature and extent of the sequences and their number or location in the Lactococcus chromosome have not been determined. Knockout insertions directed into the bacterial chromosome in the regions acquired by ul37 eliminated the AbiCinduced metam...

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Bacteriophage resistance inLactococcus

Cited by 57 publications

References 106 publications

The Pseudomonas aeruginosa generalized transducing phage φPA3 is a new member of the φKZ-like group of ‘jumbo’ phages, and infects model laboratory strains and clinical isolates from cystic fibrosis patients

The Pseudomonas aeruginosa generalized transducing phage φPA3 is a new member of the φKZ-like group of ‘jumbo’ phages, and infects model laboratory strains and clinical isolates from cystic fibrosis patients

Development of a P1 phagemid system for the delivery of DNA into Gram-negative bacteria

Genetic Analysis of Chromosomal Regions of Lactococcus lactis Acquired by Recombinant Lytic Phages

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