Acyldepsipeptide antibiotics – current state of knowledge

The objective of this paper is to review and summarize the antimicrobial efficacy of the acyldepsipeptides and to indicate the prospects of the therapeutic values of these compounds. This work is enriched by the description of the mutations within the clpP1clpP2 and c1pP3clpP4 operons of Streptomyces lividans, which are considered to be the potential mechanism of the acyldepsipeptide (ADEP)-resistance development. The researchers' conclusions demonstrated a significant impact on microorganisms including the destabilization of bacterial cell division in Bacillus subtilis 168, Staphylococcus aureus HG001 and Streptococcus pneumoniae G9A strains. The results of animal studies show higher bactericidal effectiveness of the acyldepsipeptides ADEP-2 and ADEP-4 compared to linezolid. ADEPs may be considered as a very important mechanism of defense against the increasing resistance of microorganisms . They also might prevent or reduce the risk of many epidemiological events.

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Artificial Septal Targeting of Bacillus subtilis Cell Division Proteins in Escherichia coli: an Interspecies Approach to the Study of Protein-Protein Interactions in Multiprotein Complexes

Journal of Bacteriology ◽

10.1128/jb.00462-08 ◽

2008 ◽

Vol 190 (18) ◽

pp. 6048-6059 ◽

Cited By ~ 17

Author(s):

Carine Robichon ◽

Glenn F. King ◽

Nathan W. Goehring ◽

Jon Beckwith

Keyword(s):

Escherichia Coli ◽

Bacillus Subtilis ◽

Cell Division ◽

Protein Interactions ◽

Fluorescent Protein ◽

Bacterial Cell Division ◽

Protein Protein Interactions ◽

Positive Interaction ◽

E Coli ◽

Multiprotein Complexes

ABSTRACT Bacterial cell division is mediated by a set of proteins that assemble to form a large multiprotein complex called the divisome. Recent studies in Bacillus subtilis and Escherichia coli indicate that cell division proteins are involved in multiple cooperative binding interactions, thus presenting a technical challenge to the analysis of these interactions. We report here the use of an E. coli artificial septal targeting system for examining the interactions between the B. subtilis cell division proteins DivIB, FtsL, DivIC, and PBP 2B. This technique involves the fusion of one of the proteins (the “bait”) to ZapA, an E. coli protein targeted to mid-cell, and the fusion of a second potentially interacting partner (the “prey”) to green fluorescent protein (GFP). A positive interaction between two test proteins in E. coli leads to septal localization of the GFP fusion construct, which can be detected by fluorescence microscopy. Using this system, we present evidence for two sets of strong protein-protein interactions between B. subtilis divisomal proteins in E. coli, namely, DivIC with FtsL and DivIB with PBP 2B, that are independent of other B. subtilis cell division proteins and that do not disturb the cytokinesis process in the host cell. Our studies based on the coexpression of three or four of these B. subtilis cell division proteins suggest that interactions among these four proteins are not strong enough to allow the formation of a stable four-protein complex in E. coli in contrast to previous suggestions. Finally, our results demonstrate that E. coli artificial septal targeting is an efficient and alternative approach for detecting and characterizing stable protein-protein interactions within multiprotein complexes from other microorganisms. A salient feature of our approach is that it probably only detects the strongest interactions, thus giving an indication of whether some interactions suggested by other techniques may either be considerably weaker or due to false positives.

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Asymmetric localization of the cell division machinery during Bacillus subtilis sporulation

10.1101/2020.07.22.216184 ◽

2020 ◽

Cited By ~ 1

Author(s):

Kanika Khanna ◽

Javier López-Garrido ◽

Joseph Sugie ◽

Kit Pogliano ◽

Elizabeth Villa

Keyword(s):

Bacillus Subtilis ◽

Cell Division ◽

Vegetative Growth ◽

Bacterial Cell ◽

Mother Cell ◽

Electron Tomography ◽

Leading Edge ◽

Specific Protein ◽

Bacterial Cell Division ◽

Division Plane

The mechanistic details of bacterial cell division are poorly understood. The Gram-positive bacterium Bacillus subtilis can divide via two modes. During vegetative growth, the division septum is formed at the mid cell to produce two equal daughter cells. However, during sporulation, the division septum is formed closer to one pole to yield a smaller forespore and a larger mother cell. We use cryo-electron tomography to visualize the architectural differences in the organization of FtsAZ filaments, the major orchestrators of bacterial cell division during these conditions. We demonstrate that during vegetative growth, FtsAZ filaments are present uniformly around the leading edge of the invaginating septum but during sporulation, they are only present on the mother cell side. Our data show that the sporulation septum is thinner than the vegetative septum during constriction, and that this correlates with half as many FtsZ filaments tracking the division plane during sporulation as compared to vegetative growth. We further find that a sporulation-specific protein, SpoIIE, regulates divisome localization and septal thickness during sporulation. Our data provide first evidence of asymmetric localization of the cell division machinery, and not just septum formation, to produce different cell types with diverse fates in bacteria.

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Movement Dynamics of Divisome and Penicillin-Binding Proteins (PBPs) in Cells of Streptococcus pneumoniae

10.1101/429217 ◽

2018 ◽

Author(s):

Amilcar J. Perez ◽

Yann Cesbron ◽

Sidney L. Shaw ◽

Jesus Bazan Villicana ◽

Ho-Ching T. Tsui ◽

...

Keyword(s):

Cell Division ◽

Streptococcus Pneumoniae ◽

Single Molecule ◽

Protein Complexes ◽

Bacterial Cell Division ◽

Dynamic Movement ◽

Movement Dynamics ◽

Directional Movement ◽

Tightly Coupled ◽

Filament Bundles

ABSTRACTBacterial cell division and peptidoglycan (PG) synthesis are orchestrated by the coordinated dynamic movement of essential protein complexes. Recent studies show that bidirectional treadmilling of FtsZ filaments/bundles is tightly coupled to and limiting for both septal PG synthesis and septum closure in some bacteria, but not in others. Here we report the dynamics of FtsZ movement leading to septal and equatorial ring formation in the ovoid-shaped pathogen, Streptococcus pneumoniae (Spn). Conventional and single-molecule total internal reflection fluorescence microscopy (TIRFm) showed that nascent rings of FtsZ and its anchoring and stabilizing proteins FtsA and EzrA move out from mature septal rings coincident with MapZ rings early in cell division. This mode of continuous nascent ring movement contrasts with a failsafe streaming mechanism of FtsZ/FtsA/EzrA observed in a ΔmapZ mutant and another Streptococcus species. This analysis also provides several parameters of FtsZ treadmilling in nascent and mature rings, including treadmilling velocity in wild-type cells and ftsZ(GTPase) mutants, lifetimes of FtsZ subunits in filaments and of entire FtsZ filaments/bundles, and the processivity length of treadmilling of FtsZ filament/bundles. In addition, we delineated the motion of the septal PBP2x transpeptidase and its FtsW glycosyl transferase binding partner relative to FtsZ treadmilling in Spn cells. Five lines of evidence support the conclusion that movement of the bPBP2x:FtsW complex in septa depends on PG synthesis and not on FtsZ treadmilling. Together, these results support a model in which FtsZ dynamics and associations organize and distribute septal PG synthesis, but do not control its rate in Spn.SignificanceThis study answers two long-standing questions about FtsZ dynamics and its relationship to septal PG synthesis in Spn for the first time. In previous models, FtsZ concertedly moves from midcell septa to MapZ rings that have reached the equators of daughter cells. Instead, the results presented here show that FtsZ, FtsA, and EzrA filaments/bundles move continuously out from early septa as part of MapZ rings. In addition, this study establishes that the movement of bPBP2x:FtsW complexes in septal PG synthesis depends on and likely mirrors new PG synthesis and is not correlated with the treadmilling of FtsZ filaments/bundles. These findings are consistent with a mechanism where septal FtsZ rings organize directional movement of bPBP2x:FtsW complexes dependent on PG substrate availability.

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DivIVA Is Required for Polar Growth in the MreB-Lacking Rod-Shaped Actinomycete Corynebacterium glutamicum

Journal of Bacteriology ◽

10.1128/jb.01934-07 ◽

2008 ◽

Vol 190 (9) ◽

pp. 3283-3292 ◽

Cited By ~ 94

Author(s):

Michal Letek ◽

Efrén Ordóñez ◽

José Vaquera ◽

William Margolin ◽

Klas Flärdh ◽

...

Keyword(s):

Cell Wall ◽

Bacillus Subtilis ◽

Cell Division ◽

Streptococcus Pneumoniae ◽

Corynebacterium Glutamicum ◽

Chromosome Segregation ◽

Polar Growth ◽

Cell Wall Synthesis ◽

Peptidoglycan Synthesis ◽

Polarized Cell Growth

ABSTRACT The actinomycete Corynebacterium glutamicum grows as rod-shaped cells by zonal peptidoglycan synthesis at the cell poles. In this bacterium, experimental depletion of the polar DivIVA protein (DivIVACg) resulted in the inhibition of polar growth; consequently, these cells exhibited a coccoid morphology. This result demonstrated that DivIVA is required for cell elongation and the acquisition of a rod shape. DivIVA from Streptomyces or Mycobacterium localized to the cell poles of DivIVACg-depleted C. glutamicum and restored polar peptidoglycan synthesis, in contrast to DivIVA proteins from Bacillus subtilis or Streptococcus pneumoniae, which localized at the septum of C. glutamicum. This confirmed that DivIVAs from actinomycetes are involved in polarized cell growth. DivIVACg localized at the septum after cell wall synthesis had started and the nucleoids had already segregated, suggesting that in C. glutamicum DivIVA is not involved in cell division or chromosome segregation.

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LocZ Is a New Cell Division Protein Involved in Proper Septum Placement in Streptococcus pneumoniae

mBio ◽

10.1128/mbio.01700-14 ◽

2014 ◽

Vol 6 (1) ◽

Cited By ~ 36

Author(s):

Nela Holečková ◽

Linda Doubravová ◽

Orietta Massidda ◽

Virginie Molle ◽

Karolína Buriánková ◽

...

Keyword(s):

Cell Division ◽

Streptococcus Pneumoniae ◽

Protein Kinase ◽

Bacterial Cell Division ◽

Content Type ◽

Daughter Cells ◽

Division Site ◽

Z Ring ◽

Specific Shape ◽

Cell Division Protein

ABSTRACTHow bacteria control proper septum placement at midcell, to guarantee the generation of identical daughter cells, is still largely unknown. Although different systems involved in the selection of the division site have been described in selected species, these do not appear to be widely conserved. Here, we report that LocZ (Spr0334), a newly identified cell division protein, is involved in proper septum placement inStreptococcus pneumoniae. We show thatlocZis not essential but that its deletion results in cell division defects and shape deformation, causing cells to divide asymmetrically and generate unequally sized, occasionally anucleated, daughter cells. LocZ has a unique localization profile. It arrives early at midcell, before FtsZ and FtsA, and leaves the septum early, apparently moving along with the equatorial rings that mark the future division sites. Consistently, cells lacking LocZ also show misplacement of the Z-ring, suggesting that it could act as a positive regulator to determine septum placement. LocZ was identified as a substrate of the Ser/Thr protein kinase StkP, which regulates cell division in S. pneumoniae. Interestingly, homologues of LocZ are found only in streptococci, lactococci, and enterococci, indicating that this close phylogenetically related group of bacteria evolved a specific solution to spatially regulate cell division.IMPORTANCEBacterial cell division is a highly ordered process regulated in time and space. Recently, we reported that the Ser/Thr protein kinase StkP regulates cell division in Streptococcus pneumoniae, through phosphorylation of several key proteins. Here, we characterized one of the StkP substrates, Spr0334, which we named LocZ. We show that LocZ is a new cell division protein important for proper septum placement and likely functions as a marker of the cell division site. Consistently, LocZ supports proper Z-ring positioning at midcell. LocZ is conserved only among streptococci, lactococci, and enterococci, which lack homologues of the Min and nucleoid occlusion effectors, indicating that these bacteria adapted a unique mechanism to find their middle, reflecting their specific shape and symmetry.

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Asymmetric localization of the cell division machinery during Bacillus subtilis sporulation

eLife ◽

10.7554/elife.62204 ◽

2021 ◽

Vol 10 ◽

Author(s):

Kanika Khanna ◽

Javier Lopez Garrido ◽

Joseph Sugie ◽

Kit Pogliano ◽

Elizabeth Villa

Keyword(s):

Bacillus Subtilis ◽

Cell Division ◽

Mother Cell ◽

Electron Tomography ◽

Leading Edge ◽

Specific Protein ◽

Bacterial Cell Division ◽

Daughter Cells ◽

Septal Thickness ◽

Cryo Electron Tomography

The Gram-positive bacterium Bacillus subtilis can divide via two modes. During vegetative growth, the division septum is formed at the midcell to produce two equal daughter cells. However, during sporulation, the division septum is formed closer to one pole to yield a smaller forespore and a larger mother cell. Using cryo-electron tomography, genetics and fluorescence microscopy, we found that the organization of the division machinery is different in the two septa. While FtsAZ filaments, the major orchestrators of bacterial cell division, are present uniformly around the leading edge of the invaginating vegetative septa, they are only present on the mother cell side of the invaginating sporulation septa. We provide evidence suggesting that the different distribution and number of FtsAZ filaments impact septal thickness, causing vegetative septa to be thicker than sporulation septa already during constriction. Finally, we show that a sporulation-specific protein, SpoIIE, regulates asymmetric divisome localization and septal thickness during sporulation.

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Antimicrobial Activity of Hop Resins

Journal of Food Protection ◽

10.4315/0362-028x-57.1.59 ◽

1994 ◽

Vol 57 (1) ◽

pp. 59-61 ◽

Cited By ~ 41

Author(s):

GERHARD J. HAAS ◽

RAFFI BARSOUMIAN

Keyword(s):

Escherichia Coli ◽

Staphylococcus Aureus ◽

Antimicrobial Activity ◽

Bacillus Subtilis ◽

Bacillus Megaterium ◽

Streptococcus Salivarius ◽

Resistance Development ◽

Escherichia Coli B

The antimicrobial activity of hop resins against Streptococcus salivarius, Staphylococcus aureus (two strains), Bacillus megaterium, Escherichia coli B, and Bacillus subtilis was investigated. However, resistance development was carried out on Streptococcus salivarius, Staphylococcus aureus (two strains), and Bacillus megaterium. The two hop resins used were iso-alpha resin and beta resin. Prior to resistance development, S. salivarius, S. aureus, and B. megaterium were all inhibited by the iso-alpha-hop resin in the 0.01 to 0.03% range. The beta-hop resin which, according to the literature, is more active than the iso-alpha resin initially inhibited these organisms at the 0.003 to 0.01% concentrations. The ease of resistance development varied between the different microbes, B. megaterium being the least prone to develop resistance.

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Antisense Growth Inhibition of Methicillin-Resistant Staphylococcus aureus by Locked Nucleic Acid Conjugated with Cell-Penetrating Peptide as a Novel FtsZ Inhibitor

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.03781-14 ◽

2014 ◽

Vol 59 (2) ◽

pp. 914-922 ◽

Cited By ~ 16

Author(s):

Jingru Meng ◽

Fei Da ◽

Xue Ma ◽

Ning Wang ◽

Yukun Wang ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Cell Division ◽

Nucleic Acid ◽

Bacterial Cell ◽

Locked Nucleic Acid ◽

Bacterial Cell Division ◽

Inhibitory Effects ◽

Content Type ◽

Division Process

ABSTRACTMethicillin-resistantStaphylococcus aureus(MRSA) infections are becoming increasingly difficult to treat, owing to acquired antibiotic resistance. The emergence and spread of MRSA limit therapeutic options and require new therapeutic strategies, including novel MRSA-active antibiotics. Filamentous temperature-sensitive protein Z (FtsZ) is a highly conserved bacterial tubulin homologue that is essential for controlling the bacterial cell division process in different species ofS. aureus. We conjugated a locked nucleic acid (LNA) that targetedftsZmRNA with the peptide (KFF)3K, to generate peptide-LNA (PLNA). The present study aimed to investigate whether PLNA could be used as a novel antibacterial agent. PLNA787, the most active agent synthesized, exhibited promising inhibitory effects on four pathogenicS. aureusstrainsin vitro. PLNA787 inhibited bacterial growth and resolved lethal Mu50 infections in epithelial cell cultures. PLNA787 also improved the survival rates of Mu50-infected mice and was associated with reductions of bacterial titers in several tissue types. The inhibitory effects onftsZmRNA and FtsZ protein expression and inhibition of the bacterial cell division process are considered to be the major mechanisms of PLNA. PLNA787 demonstrated activity against MRSA infectionsin vitroandin vivo. Our findings suggest thatftsZmRNA is a promising new target for developing novel antisense antibiotics.

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Bacterial SPOR domains are recruited to septal peptidoglycan by binding to glycan strands that lack stem peptides

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1508536112 ◽

2015 ◽

Vol 112 (36) ◽

pp. 11347-11352 ◽

Cited By ~ 49

Author(s):

Atsushi Yahashiri ◽

Matthew A. Jorgenson ◽

David S. Weiss

Keyword(s):

Escherichia Coli ◽

Bacillus Subtilis ◽

Cell Division ◽

Binding Site ◽

Bacterial Cell ◽

Specific Binding ◽

Bacterial Cell Division ◽

Target Proteins ◽

Division Site ◽

Lytic Transglycosylase

Bacterial SPOR domains bind peptidoglycan (PG) and are thought to target proteins to the cell division site by binding to “denuded” glycan strands that lack stem peptides, but uncertainties remain, in part because septal-specific binding has yet to be studied in a purified system. Here we show that fusions of GFP to SPOR domains from theEscherichia colicell-division proteins DamX, DedD, FtsN, and RlpA all localize to septal regions of purified PG sacculi obtained fromE.coliandBacillus subtilis. Treatment of sacculi with an amidase that removes stem peptides enhanced SPOR domain binding, whereas treatment with a lytic transglycosylase that removes denuded glycans reduced SPOR domain binding. These findings demonstrate unequivocally that SPOR domains localize by binding to septal PG, that the physiologically relevant binding site is indeed a denuded glycan, and that denuded glycans are enriched in septal PG rather than distributed uniformly around the sacculus. Accumulation of denuded glycans in the septal PG of bothE.coliandB.subtilis, organisms separated by 1 billion years of evolution, suggests that sequential removal of stem peptides followed by degradation of the glycan backbone is an ancient feature of PG turnover during bacterial cell division. Linking SPOR domain localization to the abundance of a structure (denuded glycans) present only transiently during biogenesis of septal PG provides a mechanism for coordinating the function of SPOR domain proteins with the progress of cell division.

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Movement dynamics of divisome proteins and PBP2x:FtsW in cells of Streptococcus pneumoniae

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1816018116 ◽

2019 ◽

Vol 116 (8) ◽

pp. 3211-3220 ◽

Cited By ~ 36

Author(s):

Amilcar J. Perez ◽

Yann Cesbron ◽

Sidney L. Shaw ◽

Jesus Bazan Villicana ◽

Ho-Ching T. Tsui ◽

...

Keyword(s):

Cell Division ◽

Streptococcus Pneumoniae ◽

Single Molecule ◽

Protein Complexes ◽

Bacterial Cell Division ◽

Binding Partner ◽

Dynamic Movement ◽

Movement Dynamics ◽

Tightly Coupled ◽

Filament Bundles

Bacterial cell division and peptidoglycan (PG) synthesis are orchestrated by the coordinated dynamic movement of essential protein complexes. Recent studies show that bidirectional treadmilling of FtsZ filaments/bundles is tightly coupled to and limiting for both septal PG synthesis and septum closure in some bacteria, but not in others. Here we report the dynamics of FtsZ movement leading to septal and equatorial ring formation in the ovoid-shaped pathogen, Streptococcus pneumoniae. Conventional and single-molecule total internal reflection fluorescence microscopy (TIRFm) showed that nascent rings of FtsZ and its anchoring and stabilizing proteins FtsA and EzrA move out from mature septal rings coincident with MapZ rings early in cell division. This mode of continuous nascent ring movement contrasts with a failsafe streaming mechanism of FtsZ/FtsA/EzrA observed in a ΔmapZ mutant and another Streptococcus species. This analysis also provides several parameters of FtsZ treadmilling in nascent and mature rings, including treadmilling velocity in wild-type cells and ftsZ(GTPase) mutants, lifetimes of FtsZ subunits in filaments and of entire FtsZ filaments/bundles, and the processivity length of treadmilling of FtsZ filament/bundles. In addition, we delineated the motion of the septal PBP2x transpeptidase and its FtsW glycosyl transferase-binding partner relative to FtsZ treadmilling in S. pneumoniae cells. Five lines of evidence support the conclusion that movement of the bPBP2x:FtsW complex in septa depends on PG synthesis and not on FtsZ treadmilling. Together, these results support a model in which FtsZ dynamics and associations organize and distribute septal PG synthesis, but do not control its rate in S. pneumoniae.

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