The Crystal Structure of the Lipid II-degrading Bacteriocin Syringacin M Suggests Unexpected Evolutionary Relationships between Colicin M-like Bacteriocins

Grinter, Rhys; Roszak, A.W.; Cogdell, Richard J.; Milner, Joel J.; Walker, Daniel

doi:10.1074/jbc.m112.400150

Cited by 37 publications

(62 citation statements)

References 60 publications

(75 reference statements)

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“…In addition to bacteriocins, induced prophages can also act as anticompetitor agents, allowing a strain to invade into a niche that is already colonized 7 . Pseudomonas syringae is a plant pathogen known to produce an array of antimicrobial agents, including single protein bacteriocins 8 , bacteriophage tail-derived bacteriocins 9 (termed tailocins), as well as non-proteinaceous secondary metabolites 10 . Recently there has been interest in understanding how these antimicrobials influence the ecology of this organism, as well as how they can be harnessed to control plant disease 11 .…”

Section: Introductionmentioning

confidence: 99%

Use of the Soft-agar Overlay Technique to Screen for Bacterially Produced Inhibitory Compounds

Hockett

Baltrus

2017

JoVE

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The soft-agar overlay technique was originally developed over 70 years ago and has been widely used in several areas of microbiological research, including work with bacteriophages and bacteriocins, proteinaceous antibacterial agents. This approach is relatively inexpensive, with minimal resource requirements. This technique consists of spotting supernatant from a donor strain (potentially harboring a toxic compound(s)) onto a solidified soft agar overlay that is seeded with a bacterial test strain (potentially sensitive to the toxic compound(s)). We utilized this technique to screen a library of Pseudomonas syringae strains for intraspecific killing. By combining this approach with a precipitation step and targeted gene deletions, multiple toxic compounds produced by the same strain can be differentiated. The two antagonistic agents commonly recovered using this technique are bacteriophages and bacteriocins. These two agents can be differentiated using two simple additional tests. Performing a serial dilution on a supernatant containing bacteriophage will result in individual plaques becoming less in number with greater dilution, whereas serial dilution of a supernatant containing bacteriocin will result a clearing zone that becomes uniformly more turbid with greater dilution. Additionally, a bacteriophage will produce a clearing zone when spotted onto a fresh soft agar overlay seeded with the same strain, whereas a bacteriocin will not produce a clearing zone when transferred to a fresh soft agar lawn, owing to the dilution of the bacteriocin.

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Section: Introductionmentioning

confidence: 99%

Use of the Soft-agar Overlay Technique to Screen for Bacterially Produced Inhibitory Compounds

Hockett

Baltrus

2017

JoVE

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“…During the last few years, several ColM orthologues have been identified, by sequence alignments of their C-terminal domains, in some other bacterial genera, such as Pseudomonas , Burkholderia and Pectobacterium species, and several of them have been both biochemically and structurally characterized [11,12,13,14,15]. All of the purified ColM orthologues displayed the same enzymatic activity of cleavage of lipid II and a bacteriolytic (or at least bacteriostatic) activity [11,12].…”

Section: Introductionmentioning

confidence: 99%

“…All of the purified ColM orthologues displayed the same enzymatic activity of cleavage of lipid II and a bacteriolytic (or at least bacteriostatic) activity [11,12]. Yet, despite their similarities in terms of enzymatic activity, these orthologues did not share the same 3D structure, especially in their N-terminal domains.…”

Section: Introductionmentioning

confidence: 99%

“…Yet, despite their similarities in terms of enzymatic activity, these orthologues did not share the same 3D structure, especially in their N-terminal domains. On the one hand, ColM shared a very compact structure with PaeM and syringacin M (also named PsyM), i.e., its orthologues from Pseudomonas aeruginosa and Pseudomonas syringae , respectively, where the usual three structural domains did not form distinct entities [12,13]. On the other hand, the structure of pectocin M2 from Pectobacterium carotovorum and probably also that of pectocin M1, since it has been strongly suggested that both pectocins M1 and M2 used the same outer membrane receptor on Pectobacterium spp.…”

Section: Introductionmentioning

confidence: 99%

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Pectocin M1 (PcaM1) Inhibits Escherichia coli Cell Growth and Peptidoglycan Biosynthesis through Periplasmic Expression

et al. 2016

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Colicins are bacterial toxins produced by some Escherichia coli strains. They exhibit either enzymatic or pore-forming activity towards a very limited number of bacterial species, due to the high specificity of their reception and translocation systems. Yet, we succeeded in making the colicin M homologue from Pectobacterium carotovorum, pectocin M1 (PcaM1), capable of inhibiting E. coli cell growth by bypassing these reception and translocation steps. This goal was achieved through periplasmic expression of this pectocin. Indeed, when appropriately addressed to the periplasm of E. coli, this pectocin could exert its deleterious effects, i.e., the enzymatic degradation of the peptidoglycan lipid II precursor, which resulted in the arrest of the biosynthesis of this essential cell wall polymer, dramatic morphological changes and, ultimately, cell lysis. This result leads to the conclusion that colicin M and its various orthologues constitute powerful antibacterial molecules able to kill any kind of bacterium, once they can reach their lipid II target. They thus have to be seriously considered as promising alternatives to antibiotics.

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“…During our characterization of the pectocins, it became apparent that in addition to being susceptible to a ferredoxin-containing bacteriocin, Pectobacterium spp. are also able to utilize plant ferredoxins as an iron source under iron-limiting conditions14. Moreover, competition experiments showed that both the pectocins and ferredoxin are bound by the same receptor during cell entry15.…”

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confidence: 99%

Structure of the bacterial plant-ferredoxin receptor FusA

et al. 2016

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Iron is a limiting nutrient in bacterial infection putting it at the centre of an evolutionary arms race between host and pathogen. Gram-negative bacteria utilize TonB-dependent outer membrane receptors to obtain iron during infection. These receptors acquire iron either in concert with soluble iron-scavenging siderophores or through direct interaction and extraction from host proteins. Characterization of these receptors provides invaluable insight into pathogenesis. However, only a subset of virulence-related TonB-dependent receptors have been currently described. Here we report the discovery of FusA, a new class of TonB-dependent receptor, which is utilized by phytopathogenic Pectobacterium spp. to obtain iron from plant ferredoxin. Through the crystal structure of FusA we show that binding of ferredoxin occurs through specialized extracellular loops that form extensive interactions with ferredoxin. The function of FusA and the presence of homologues in clinically important pathogens suggests that small iron-containing proteins represent an iron source for bacterial pathogens.

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The Crystal Structure of the Lipid II-degrading Bacteriocin Syringacin M Suggests Unexpected Evolutionary Relationships between Colicin M-like Bacteriocins

Cited by 37 publications

References 60 publications

Use of the Soft-agar Overlay Technique to Screen for Bacterially Produced Inhibitory Compounds

Use of the Soft-agar Overlay Technique to Screen for Bacterially Produced Inhibitory Compounds

Pectocin M1 (PcaM1) Inhibits Escherichia coli Cell Growth and Peptidoglycan Biosynthesis through Periplasmic Expression

Structure of the bacterial plant-ferredoxin receptor FusA

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