The lysis cassette of bacteriophage фKMV encodes a signal-arrest-release endolysin and a pinholin

Briers, Yves; Peeters, Liesbet M.; Volckaert, Guido; Lavigne, Rob

doi:10.4161/bact.1.1.14868

Cited by 46 publications

(61 citation statements)

References 23 publications

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“…E. coli bacteriophage P1 lysin, for example, showed the presence of N-terminal SAR sequence when its periplasmic and cytoplasmic fractions were tested on SDS-PAGE, suggesting that the SAR sequence remains attached to the endolysin even after export (Xu et al, 2004). Similar observations were made for Erwinia amylovora phage ERA103 and other bacteriophages (Kuty et al, 2010;Briers et al, 2011).…”

Section: Regulation During Translocationsupporting

confidence: 67%

“…Although the study verifies the requirement of the disruption of proton motive force in endolysin activation, a direct involvement of fOg44 holin in the phage's endolysin activation has currently not been examined. In another report, the expression of Pseudomonas aeruginosa phage w KMV endolysin resulted in the lysis of E. coli 60-90 min after induction, whereas the co-expression of endolysin and holin led to bacterial cell lysis within 15 min of induction (Briers et al, 2011).…”

Section: Regulation By Inactivationmentioning

confidence: 99%

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Insights into the regulation of bacteriophage endolysin: multiple means to the same end

Pohane¹,

Jain²

2015

Microbiology

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Antibiotics are the molecules of choice to treat bacterial infections. However, because of the rapid emergence of drug-resistant bacteria, alternative modes of combating infections are being envisaged. Bacteriophages, which infect and lyse bacterial cells, may function as effective antimicrobial agents. Most bacteriophages produce their own peptidoglycan hydrolase called endolysin or lysin, which breaks down the cell wall of bacteria and aids in the release of newly assembled virions. Here, we discuss several findings that help us in understanding how endolysins are regulated. We observe that there is no common mechanism that is followed in all cases. Many different modes of activity regulation have been observed in endolysins, including regulation of protein expression, translocation across the cell membrane and post-translational modifications. These processes not only demonstrate how endolysins are made dependent on other accessory proteins and non-protein factors for their synthesis, translocation across the cytoplasmic membrane and activity, but also show how autoregulation helps in maintaining the enzyme in an inactive form. Various regulatory mechanisms that are discussed are particularly applicable to endolysins. Nevertheless, a detailed study of these methods opens new avenues of investigation in the area of protein translocation systems and the novel ways of enzyme activation and regulation in bacteria.

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Section: Regulation During Translocationsupporting

confidence: 67%

Section: Regulation By Inactivationmentioning

confidence: 99%

Insights into the regulation of bacteriophage endolysin: multiple means to the same end

Pohane¹,

Jain²

2015

Microbiology

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“…Another alternative to the lambda paradigm is the SAR system, in which the endolysin possesses a noncleavable N-terminal type II signal anchor that remains part of the mature endolysin and stays embedded in the inner cell membrane in an inactive form (42,92,93). Consequently, these endolysins require pinholins to cause a membrane depolarization to release the SAR anchor enabling the endolysin to get access to the cell wall (92,94,95).…”

Section: Resultsmentioning

confidence: 99%

Molecular Aspects and Comparative Genomics of Bacteriophage Endolysins

Oliveira

Melo

Santos

et al. 2013

J Virol

241

247

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Phages are recognized as the most abundant and diverse entities on the planet. Their diversity is determined predominantly by their dynamic adaptation capacities when confronted with different selective pressures in an endless cycle of coevolution with a widespread group of bacterial hosts. At the end of the infection cycle, progeny virions are confronted with a rigid cell wall that hinders their release into the environment and the opportunity to start a new infection cycle. Consequently, phages encode hydrolytic enzymes, called endolysins, to digest the peptidoglycan. In this work, we bring to light all phage endolysins found in completely sequenced double-stranded nucleic acid phage genomes and uncover clues that explain the phage-endolysin-host ecology that led phages to recruit unique and specialized endolysins.

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“…TMD1 (residues 13 to 29) is not predicted by TMHMM and has characteristics of a SAR domain, with a high percentage of weakly hydrophobic or polar residues. The class II topology and the presence of a SAR domain followed by a typical TMD suggest that gp45 is a pinholin, similar to phiKMV gp45 (47). Prado gene 49 encodes an endolysin, based on homology to other phiKMV-like phage endolysins in the database.…”

Section: Resultsmentioning

confidence: 99%

“…The lysis cassette of phage phiKMV has been studied in detail and is organized with genes encoding holin, endolysin, i-spanin (formerly Rz), and o-spanin (formerly Rz1) immediately downstream of TerS and TerL (47). The lysis cassette of Prado is similar to that of phiKMV, except that the predicted holin is found upstream of the endolysin, separated by three genes, encoding TerS (gene 46), TerL (gene 47), and a hypothetical protein (gene 48) (Fig.…”

Section: Resultsmentioning

confidence: 99%

Characterization of Novel Virulent Broad-Host-Range Phages of Xylella fastidiosa and Xanthomonas

Ahern

Das

Bhowmick

et al. 2013

Journal of Bacteriology

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The xylem-limited bacterium Xylella fastidiosa is the causal agent of several plant diseases, most notably Pierce's disease of grape and citrus variegated chlorosis. We report the isolation and characterization of the first virulent phages for X. fastidiosa, siphophages Sano and Salvo and podophages Prado and Paz, with a host range that includes Xanthomonas spp. Phages propagated on homologous hosts had observed adsorption rate constants of ϳ4 ؋ 10 ؊12 ml cell ؊1 min ؊1 for X. fastidiosa strain Temecula 1 and ϳ5 ؋ 10 ؊10 to 7 ؋ 10 ؊10 ml cell ؊1 min ؊1 for Xanthomonas strain EC-12. Sano and Salvo exhibit >80% nucleotide identity to each other in aligned regions and are syntenic to phage BcepNazgul. We propose that phage BcepNazgul is the founding member of a novel phage type, to which Sano and Salvo belong. The lysis genes of the Nazgul-like phage type include a gene that encodes an outer membrane lipoprotein endolysin and also spanin gene families that provide insight into the evolution of the lysis pathway for phages of Gram-negative hosts. Prado and Paz, although exhibiting no significant DNA homology to each other, are new members of the phiKMV-like phage type, based on the position of the single-subunit RNA polymerase gene. The four phages are type IV pilus dependent for infection of both X. fastidiosa and Xanthomonas. The phages may be useful as agents for an effective and environmentally responsible strategy for the control of diseases caused by X. fastidiosa.T he plant-pathogenic bacterium Xylella fastidiosa is the causative agent of a number of economically important diseases, including Pierce's disease of grape, phony peach disease, periwinkle wilt, citrus variegated chlorosis, almond leaf scorch, oleander leaf scorch, and coffee leaf scorch (1). Existing disease control methods are often only partially successful and, in the case of systemic insecticides used to control vector populations, may be potentially harmful to the environment (2, 3). Recently, there has been renewed interest in the application of bacteriophages (phage) as an environmentally acceptable mode for the control of bacterial plant disease (4).To this end, our group isolated and characterized the first temperate (lysogenic) phage of X. fastidiosa, Xfas53 (5). However, temperate phage should not be used as biocontrol agents because of their capacity for lysogenic conversion, superinfection immunity, and risk of generalized transduction. Instead, virulent (lytic) phages are needed for an effective and sustainable phage-based control strategy (6). Here we report the isolation and characterization of the first virulent siphophages and podophages for X. fastidiosa that are members of the Nazgul-like phage and the phiKMV-like phage types, respectively. MATERIALS AND METHODSBacterial strains and culture conditions. Bacterial strains and plasmids used in this study are listed in Table 1. X. fastidiosa strains were cultured at 28°C in PW-M broth (PW-MB) (7) or on PW-M agar (PW-MA) plates (PW-MB with 20 g/liter plant cell culture-tested agar [...

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The lysis cassette of bacteriophage фKMV encodes a signal-arrest-release endolysin and a pinholin

Cited by 46 publications

References 23 publications

Insights into the regulation of bacteriophage endolysin: multiple means to the same end

Insights into the regulation of bacteriophage endolysin: multiple means to the same end

Molecular Aspects and Comparative Genomics of Bacteriophage Endolysins

Characterization of Novel Virulent Broad-Host-Range Phages of Xylella fastidiosa and Xanthomonas

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