Fluorescent D-amino-acids reveal bi-cellular cell wall modifications important for Bdellovibrio bacteriovorus predation

Kuru, Erkin; Lambert, Carey; Rittichier, Jonathan; Till, Rob; Ducret, Adrien; Derouaux, Adeline; Gray, Joe; Biboy, Jacob; Vollmer, Waldemar; VanNieuwenhze, Michael S.; Brun, Yves V.; Sockett, R. Elizabeth

doi:10.1038/s41564-017-0029-y

Cited by 108 publications

(147 citation statements)

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“…2E). During the late stage of B. bacteriovorus cell growth, the filament can reach a length exceeding the bdelloplast diameter and begin to curve and overlap itself (13, 15, 36). In such cases, it becomes difficult to analyze the filament length and replisome number.…”

Section: Resultsmentioning

confidence: 99%

“…Although the cell biology of B. bacteriovorus has gained considerable recent interest (13–15) we know very little about the dynamics of chromosome replication in this predatory bacterium. B. bacteriovorus possesses a single circular chromosome (16) that contains all essential genes (e.g., those encoding the Dna proteins) and elements (e.g., an origin of chromosomal replication, oriC ) required for its own replication.…”

Section: Introductionmentioning

confidence: 99%

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Dynamics of chromosome replication and its relationship to predatory attack lifestyles in Bdellovibrio bacteriovorus

Makowski

Trojanowski

Till

et al. 2019

Preprint

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13Bdellovibrio bacteriovorus is a small Gram-negative, an obligate predatory bacterium that is 14 largely found in wet, aerobic environments (i.e. soil). This bacterium attacks and invades other 15 Gram-negative bacteria, including animal and plant pathogens. The intriguing life cycle of B. 16 bacteriovorus consists of two phases: a free-living non-replicative attack phase wherein the 17 predatory bacterium searches for its prey, and a reproductive phase, in which B. bacteriovorus 18 degrades a host's macromolecules and reuses them for its own growth and chromosome 19 replication. Although the cell biology of this predatory bacterium has gained considerable 20 interest in recent years, we know almost nothing about the dynamics of chromosome replication 21 in B. bacteriovorus. Here, we performed a real-time investigation into the subcellular 22 localization of the replisome(s) in single cells of B. bacteriovorus. Our results confirm that in 23 B. bacteriovorus chromosome replication fires only during the reproductive phase, and show 24 for the first time that this predatory bacterium exhibits a novel spatiotemporal arrangement of 25 chromosome replication. The replication process starts at the invasive pole of the predatory 26 bacterium inside the prey cell and proceeds until several copies of the chromosome have been 27 completely synthesized. This chromosome replication is not coincident with the predator-cell 28 division, and it terminates shortly before synchronous predator-filament septation occurs. In 29 addition, we demonstrate that if this lifecycle fails in some cells of B. bacteriovorus, they can 30 instead use two prey cells sequentially to complete their life cycle. 31 Importance 32 New strategies are needed to combat multidrug-resistant bacterial infections. Application of the 33 predatory bacterium, Bdellovibrio bacteriovorus, which kills other bacteria including 34 pathogens, is considered promising for bacterial infections. The B. bacteriovorus life cycle 35 consists of two phases, a free-living, invasive attack phase and an intracellular reproductive 36 phase, in which this predatory bacterium degrades the host's macromolecules and reuses them 37 3 for its own growth. To understand the use of B. bacteriovorus as a 'living antibiotic', it is first 38 necessary to dissect its life cycle including chromosome replication. Here, we present for the 39 first time a real-time investigation into subcellular localization of chromosome replication in a 40 single cells of B. bacteriovorus. This process initiates at the invasion pole of B. bacteriovorus 41 and proceeds until several copies of the chromosome have been completely synthesized. 42 Interestingly, we demonstrate that some cells of B. bacteriovorus require two prey cells 43 sequentially to complete their life cycle. 44 45Bdellovibrio bacteriovorus is a small (0.2-to 0.5-μm wide and 0.5-to 2.5-μm long) 46 Gram-negative bacterium that is unusual in its ability to invade and kill other Gram-negative 47 bacteria. Moreover, it was demonstrated th...

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Dynamics of chromosome replication and its relationship to predatory attack lifestyles in Bdellovibrio bacteriovorus

Makowski

Trojanowski

Till

et al. 2019

Preprint

Self Cite

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“…1C, S2C,D, S3). Thus, like Bdellovibrio (6,8), FDAA incorporation in Msm is primarily LDTdependent. LDTs can also exchange non-canonical D-amino acids onto PG tetrapeptides in Vibrio cholera (9).…”

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confidence: 99%

Maturing Mycobacterial Peptidoglycan Requires Non-canonical Crosslinks to Maintain Shape

Baranowski

Sham

et al. 2018

Preprint

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In most well-studied rod-shaped bacteria, peptidoglycan is primarily crosslinked by penicillin binding proteins (PBPs). However, in mycobacteria L,D-transpeptidase (LDT)mediated crosslinks are highly abundant. To elucidate the role of these unusual crosslinks, we characterized mycobacterial cells lacking all LDTs. We find that LDT-mediated crosslinks are required for rod shape maintenance specifically at sites of aging cell wall, a byproduct of polar elongation. Asymmetric polar growth leads to a non-uniform distribution of these two types of crosslinks in a single cell. Consequently, in the absence of LDT-mediated crosslinks, PBPcatalyzed crosslinks become more important. Because of this, Mycobacterium tuberculosis is more rapidly killed using a combination of drugs capable of PBP and LDT inhibition. Thus, knowledge about the single-cell distribution of drug targets can be exploited to more effectively treat this pathogen.One Sentence Summary: Polar elongating mycobacteria utilize specific cell wall chemistry to maintain rod shape at sites of aging cell wall. Main Text:Peptidoglycan (PG) is an essential component of all bacterial cells (1), and the target of many antibiotics. PG consists of linear glycan strands crosslinked by short peptides to form a continuous molecular cage surrounding the plasma membrane. This structure maintains cell shape and protects the plasma membrane from rupture. Our understanding of PG is largely

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“…In particular, four HD100 protein-coding genes that are missing in NC01 are predicted to play a role in peptidoglycan metabolism. Invasion of a prey cell by Bdellovibrio involves opening a hole in the prey cell peptidoglycan, then resealing and reshaping peptidoglycan to form a rounded bdelloplast (11, 15). If NC01 is missing enzymes that play a role in these processes, it may impact the ability of this strain to invade prey cells and establish itself in the prey periplasm.…”

Section: Discussionmentioning

confidence: 99%

“…During the attack phase, small, highly motile Bdellovibrio cells seek out and attach to Gram-negative prey bacteria. Bdellovibrio then invades the prey periplasm by using lytic enzymes to open a small hole in the prey outer membrane and cell wall (11). Both attachment and invasion may be mediated by Type IV pili expressed by Bdellovibrio (12–14).…”

Section: Introductionmentioning

confidence: 99%

Variation in genome content and predatory phenotypes betweenBdellovibriosp. NC01 isolated from soil andB. bacteriovorustype strain HD100

Williams

Cullen

DeGiorgis

et al. 2019

Preprint

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13Defining phenotypic and associated genotypic variation among Bdellovibrio may further 14 our understanding of how this genus attacks and kills different Gram-negative bacteria. We 15 isolated Bdellovibrio sp. NC01 from soil. Analysis of 16S rRNA gene sequences and average 16 amino acid identity showed that NC01 belongs to a different species than the type species 17 bacteriovorus. By clustering amino acid sequences from completely sequenced Bdellovibrio and 18 comparing the resulting ortholog groups to a previously published analysis, we defined a "core 19 genome" of 778 protein-coding genes and identified four protein-coding genes that appeared to 20 be missing only in NC01. To determine how horizontal gene transfer (HGT) may have impacted 21 NC01 genome evolution, we performed genome-wide comparisons of Bdellovibrio nucleotide 22 sequences, which indicated that eight NC01 genomic regions were likely acquired by HGT. To 23 investigate how genome variation may impact predation, we compared protein-coding gene 24 content between NC01 and the B. bacteriovorus type strain HD100, focusing on genes 25 implicated as important in successful killing of prey. Of these, NC01 is missing ten genes that 26 may play roles in lytic activity during predation. Compared to HD100, NC01 kills fewer tested 27 prey strains and kills E. coli ML35 less efficiently. NC01 causes a smaller log reduction in 28 ML35, after which the prey population recovers and the NC01 population decreases. In addition, 29 NC01 forms turbid plaques on lawns of E. coli ML35, in contrast to clear plaques formed by 30 HD100. Linking phenotypic variation in interactions between Bdellovibrio and Gram-negative 31 bacteria with underlying Bdellovibrio genome variation is valuable for understanding the 32 ecological significance of predatory bacteria and evaluating their effectiveness in clinical 33 applications.34 35Since their discovery in the early 1960s, predatory bacteria have been isolated from a 36 wide range of terrestrial and aquatic environments, including some with extreme conditions such 37 as high temperature or salinity (1). The impact of predatory bacteria in these environments is not 38 fully known, but recent studies indicate they may play an important role in shaping microbial 39 communities (2-4). In addition, their ability to attack and kill Gram-negative bacterial pathogens 40 has generated interest in developing predatory bacteria as biocontrol agents to combat drug-41 resistant infections (5-7). Given their potential ecological significance and possible clinical 42 applications, it is important to define phenotypic and genotypic variation among predatory 43 bacteria. In particular, variation in predatory phenotypes such as prey range and predation 44 efficiency will affect how different strains of predatory bacteria impact microbial communities, 45 and this phenotypic variation may be determined by variation in genome content. To further our 46 understanding of phenotypic and genome variation in predatory bacteria, we isolated, assayed 4...

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Fluorescent D-amino-acids reveal bi-cellular cell wall modifications important for Bdellovibrio bacteriovorus predation

Cited by 108 publications

References 43 publications

Dynamics of chromosome replication and its relationship to predatory attack lifestyles in Bdellovibrio bacteriovorus

Dynamics of chromosome replication and its relationship to predatory attack lifestyles in Bdellovibrio bacteriovorus

Maturing Mycobacterial Peptidoglycan Requires Non-canonical Crosslinks to Maintain Shape

Variation in genome content and predatory phenotypes betweenBdellovibriosp. NC01 isolated from soil andB. bacteriovorustype strain HD100

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