Characterization of Two Virulent Phages of Lactobacillus plantarum

Marcó, Mariángeles Briggiler; Garneau, Josiane E; Tremblay, Denise M.; Quiberoni, Andrea del Luján; Moineau, Sylvain

doi:10.1128/aem.02565-12

Cited by 38 publications

(34 citation statements)

References 70 publications

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“…29 In phage B2, only 2 of the 6 tRNA genes corresponded to codons more frequently used by the phage than its Lactobacillus plantarum host. 30 Limor-Waisberg et al looked at cyanophages with very different genome size and GC content: T7-like podoviruses have small genomes, few tRNA genes and only infect hosts with similar GC content, while T4-like myoviruses have larger genomes, more tRNA genes, and can infect hosts that differ in GC content. 21 Among the myoviruses, greater difference in GC content between host and phages was associated with more tRNA genes in these phages.…”

Section: Codon Versus Amino Acid Usagementioning

confidence: 99%

Testing hypotheses for the presence of tRNA genes in mycobacteriophage genomes

et al. 2016

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The presence of tRNA genes in bacteriophages has been explained on the basis of codon usage (tRNA genes are retained in the phage genome if they correspond to codons more common in the phage than in its host) or amino acid usage (independent of codon, the amino acid corresponding to the retained tRNA gene is more common in the phage genome than in the bacterial host). The existence of a large database of sequenced mycobacteriophages, isolated on the common host Mycobacterium smegmatis, allows us to test the above hypotheses as well as explore other hypotheses for the presence of tRNA genes. Our analyses suggest that amino acid rather than codon usage better explains the presence of tRNA genes in mycobacteriophages. However, closely related phages that differ in the presence of tRNA genes in their genomes are capable of lysing the common bacterial host and do not differ in codon or amino acid usage. This suggests that the benefits of having tRNA genes may be associated with either growth in the host or the ability to infect more hosts (i.e., host range) rather than simply infecting a particular host.

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Section: Codon Versus Amino Acid Usagementioning

confidence: 99%

Testing hypotheses for the presence of tRNA genes in mycobacteriophage genomes

et al. 2016

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“…In fact, among phages that infect Lactobacillus species, the Ldl1 tail length is second only to that of L. plantarum phage B2, which was isolated in 1971 and which was originally reported to display a tail length of 500 nm (33). However, a recent study found the tail length of this phage to be 240 Ϯ 3 nm, with no reasons being given for the discrepancy (34). Also evident is a minihead variant of Ldl1, exhibiting a much smaller head with the long tail still present.…”

Section: Resultsmentioning

confidence: 83%

“…The latter two proteins in turn exhibit similarity to a DNA polymerase III protein (␣ subunit) from Bacillus phage SPBc2 (34). ORF37 Ldl1 also shows 41% identity to a putative DNA polymerase of Bacillus subtilis subsp.…”

Section: Resultsmentioning

confidence: 98%

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Novel Phage Group Infecting Lactobacillus delbrueckii subsp. lactis, as Revealed by Genomic and Proteomic Analysis of Bacteriophage Ldl1

Casey

Mahony

Neve

et al. 2015

Appl Environ Microbiol

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e Ldl1 is a virulent phage infecting the dairy starter Lactobacillus delbrueckii subsp. lactis LdlS. Electron microscopy analysis revealed that this phage exhibits a large head and a long tail and bears little resemblance to other characterized phages infecting Lactobacillus delbrueckii. In vitro propagation of this phage revealed a latent period of 30 to 40 min and a burst size of 59.9 ؎ 1.9 phage particles. Comparative genomic and proteomic analyses showed remarkable similarity between the genome of Ldl1 and that of Lactobacillus plantarum phage ATCC 8014-B2. The genomic and proteomic characteristics of Ldl1 demonstrate that this phage does not belong to any of the four previously recognized L. delbrueckii phage groups, necessitating the creation of a new group, called group e, thus adding to the knowledge on the diversity of phages targeting strains of this industrially important lactic acid bacterial species. L actobacillus delbrueckii subsp. lactis is a member of the lactic acid bacteria (LAB) and is commonly used in the production of commercial fermented milk products, such as Emmental-like cheeses, where it is employed as a starter culture contributing to the acidification and the organoleptic properties of the final product (1, 2). Bacteriophage (or phage) infection of these starter strains represents a major hurdle to their technological activity, as this may cause lysis of the starter, which in turn may lead to reduced acidification activity and a poor(er)-quality product, with a consequent negative economic impact (3).The commercial importance of L. delbrueckii fermentations has catalyzed extensive research into the occurrence, diversity, and impact of its infecting phages. Currently, L. delbrueckii phages are organized into four distinct groups (designated groups a, b, c, and d), classified based on DNA homology by hybridization (4-6) and, more recently, by comparative genome analysis (7). L. delbrueckii phages belonging to groups a and c have enjoyed considerable scientific scrutiny, with phage LL-H representing a prototype Lactobacillus phage. LL-H was originally isolated in 1972 (8), and its full genome sequence and transcriptional map, which revealed two distinct phases of transcription, have been determined (9). Furthermore, research on phage-host interactions, culminating in the identification of gp71 as the receptor-binding protein of LL-H (10) and lipoteichoic acids as the recognized receptor, have significantly advanced our understanding of this phage and the way in which it recognizes its host (11, 12). Also, the prolate-head, temperate phage JCL1032, being a member of group c L. delbrueckii phages, has been subjected to considerable scientific characterization, including genome sequencing (13), analysis of genomic integration (14), and identification of lipoteichoic acid as its receptor (12).Due to their apparent increased frequency of isolation, research has recently focused on group b phages, facilitated by the sequencing of six phages, c5 and LL-Ku (13); Ld3, Ld17, and Ld25A (7); and ph...

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“…1B and C). This suggested that the major capsid protein was likely processed (27). The ORF10 product (MTP) was also found in two protein bands (bands 8 and 9) with molecular masses of 26 and 22 kDa, respectively.…”

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confidence: 89%

Characterization of a Novel Panton-Valentine Leukocidin (PVL)-Encoding Staphylococcal Phage and Its Naturally PVL-Lacking Variant

Haddad

Moineau

2013

Appl Environ Microbiol

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A new siphophage (LH1) was isolated from raw milk using a Staphylococcus aureus ST352 host. Its genome (46,048 bp, 57 open reading frames) includes the two genes encoding Panton-Valentine leukocidin (PVL), a virulence factor usually harbored by S. aureus prophages. Nine structural proteins were identified, including a tail protein generated through a ؉1 frameshift. A phage lytic mutant was isolated, and its analysis revealed the deletion of genes coding for the PVL and an integrase. The deletion likely occurred through recombination between direct repeats. Staphylococcus aureus is a bacterial pathogen that can infect humans, animals, and plants and may contaminate foods. Some strains harbor several pathogenic and virulent components, including exotoxins, such as enterotoxins (SE), exfoliatins, toxic shock syndrome toxin (TSST), hemolysins, and Panton-Valentine leukocidin (PVL) (1, 2). The genes coding for this toxin are located on prophages (3). The PVL toxin belongs to the synergohymenotropic toxin family and is composed of two subunits, named LukF-PV and LukS-PV (4-6). These secretory proteins act together by forming pores in cell membranes and lysing neutrophils and macrophages, thus leading to pneumonia and eventual cell death (7,8).Since the discovery of the PVL toxin by Panton and Valentine in 1932 (5), at least eight PVL-encoding phages belonging to the Siphoviridae family have been characterized (3, 9-11). They can be classified into three different groups according to the replication/ transcription region and morphogenesis module as well as their host range (9,11,12). Members of groups 1 and 3 have isometric capsids, and members of group 2 have elongated capsids. The presence of the same toxin genes in morphologically distinct phages suggests that they can be readily exchanged, possibly during coinfection.Unlike temperate phages, virulent phages can be used as biocontrol agents against pathogenic strains in medical and food applications. The presence of an integrase gene and toxin genes in phage genomes leads to poor antibacterial efficacy and safety concerns, respectively. Consequently, only virulent phages (or temperate phages replicating on indicator strains) lacking any virulence factor have been used as biocontrol agents against S. aureus in foods (13-16) and in animal models (17)(18)(19). In an effort to isolate new virulent phages to control Staphylococcus contaminations in dairy products, we analyzed raw milk samples for the presence of staphylococcal phages.One raw milk sample was obtained from each of six dairy farms. One hundred microliters of an overnight culture of S. aureus 01 (ST352) grown in tryptic soy broth (TSB) at 37°C was added to 5 ml of 2ϫ TSB and 5 ml of the milk sample. The culture was incubated overnight at 37°C. Five milliliters of the first amplification was added to 5 ml of 2ϫ TSB, as well as 100 l of the overnight bacterial culture, and incubated overnight at 37°C. The latter step was repeated once. Then the presence of phages was tested by depositing 5 l of the last amplif...

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Characterization of Two Virulent Phages of Lactobacillus plantarum

Cited by 38 publications

References 70 publications

Testing hypotheses for the presence of tRNA genes in mycobacteriophage genomes

Testing hypotheses for the presence of tRNA genes in mycobacteriophage genomes

Novel Phage Group Infecting Lactobacillus delbrueckii subsp. lactis, as Revealed by Genomic and Proteomic Analysis of Bacteriophage Ldl1

Characterization of a Novel Panton-Valentine Leukocidin (PVL)-Encoding Staphylococcal Phage and Its Naturally PVL-Lacking Variant

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