Targeting cell division: Small-molecule inhibitors of FtsZ GTPase perturb cytokinetic ring assembly and induce bacterial lethality

Margalit, Danielle N.; Romberg, Laura; Mets, Rebecca B.; Hebert, Alan M.; Mitchison, Timothy J.; Kirschner, Marc W.; RayChaudhuri, Debabrata

doi:10.1073/pnas.0404439101

Cited by 195 publications

(206 citation statements)

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“…Inhibitors of FtsZ have been reported. [26][27][28][29][30][31] Edeine B 1 is a potentially useful tool for studying the mechanism of bacterial cell division.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of Septation in Bacillus subtilis by a Peptide Antibiotic, Edeine B1

Shimotohno

Kawamura

Natori

et al. 2010

Biological & Pharmaceutical Bulletin

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The antibiotics, edeine A 1 and B 1 , are well-known inhibitors of protein biosynthesis in both prokaryote and eukaryote cells. Edeine A 1 and B 1 show almost the same biological activity. We show herein that edeine B 1 exhibits a lethal action in Bacillus subtilis, inhibiting septation via a mechanism different from the inhibition of protein synthesis by this antibiotic.The complex process of cell division is closely interconnected with other cellular processes. Chromosomal DNA replication, nuclear division, and cell division must be coordinated in time and space. DNA metabolism plays a pivotal role in the cell division events controlled by the partitioning of newly replicated chromosomes to daughter cells. This involves chromosomal DNA replication, the positioning of a septum at the midpoint of the cell, the assembly of cytoskeletal elements, and the cleavage of the septum after chromosome segregation. Genetic studies have unraveled the process of cell division by the analysis of filamenting temperaturesensitive mutants. In prokaryotes, filamenting temperaturesensitive (fts) genes produce the proteins essential for cell division. Known fts mutants include ftsA, ftsI, ftsK, ftsL, ftsN, ftsW and ftsZ. [1][2][3][4][5][6][7][8] Inhibitors of protein synthesis exhibit bacteriostatic action except for the aminoglycoside (AG) antibiotics, 9) suggesting that the inhibition of protein synthesis by AG antibiotics is not directly involved in their bactericidal action. The bactericidal action of AG antibiotic is assumed to be based on their inhibition of DNA replication. AG antibiotic blocks the initiation of DNA replication, 10) and interrupts oriC-membrane attachment. 11) The oriC-membrane attachment [12][13][14] is crucial for subsequent initiation of DNA replication, chromosome segregation, and septation of bacterial cells. Blocking DNA replication is known to lead to the save our ships (SOS) response in Escherichia coli, thereby inhibiting cell division. 15) We demonstrate herein that edeine B 1 inhibits cell division, exerting its bactericidal action by inhibiting FtsZ assembly, independently of its inhibitory effect on protein synthesis. MATERIALS AND METHODSPurification of Edeine B 1 Edeine B 1 was isolated from a culture filtrate of Bacillus brevis TTO2-8. The purified edeine B 1 yielded a single TLC spot, a single HPLC peak and the predicted structure based on NMR and FAB-MAS analysis. Edeine B 1 is a linear basic oligopeptide antibiotic consisting of five amino acid residues (isotyrosine, isoserine, a,b-diaminopropionic acid, 2,6-diamino-7-hydroxyazelanic acid, and glycine) and an organic base (guanylspermidine) produced by B. brevis. 16,17) Bacterial Strains B. subtilis 168 and mutant FtsZ ts1 (a temperature sensitive mutant of B. subtilis 168) were used.Viable Cell Number The viable cell number was counted by plating on LB (Luria-Bertani) agar, and recording the number of colonies that developed after 24 h incubation.Western Blotting Anti-FtsZ rabbit antibody raised against a chemically synthesized carboxyl t...

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“…Inhibitors of FtsZ have been reported. [26][27][28][29][30][31] Edeine B 1 is a potentially useful tool for studying the mechanism of bacterial cell division.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of Septation in Bacillus subtilis by a Peptide Antibiotic, Edeine B1

Shimotohno

Kawamura

Natori

et al. 2010

Biological & Pharmaceutical Bulletin

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“…Nevertheless, thiostrepton for IF-2, pulvomycin and GE2270A for EF-Tu, and PC190723, Berberine and Zantrins for FtsZ have been exploited and investigated for their inhibitory activity towards those GTPases. [29][30][31][32][33][34][35][36][37][38] The recent technological advances in computational modeling of protein structures have a crucial role in drug design and its retrieval. In our study, structure-based pharmacophore design screened a relatively small number of candidate molecules and we successfully identified three inhibitors (SBI-34462, -34566, and -34612) for an essential bacterial GTPase Der that has been unexploited for antibacterial target.…”

Section: Models Of Der Gtpase Complexes With Bioactive Compoundsmentioning

confidence: 99%

Structure-based design and screening of inhibitors for an essential bacterial GTPase, Der

Hwang

Tseitin²,

Ramnarayan³

et al. 2012

J Antibiot

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Der is an essential and widely conserved GTPase that assists assembly of a large ribosomal subunit in bacteria. Der associates specifically with the 50S subunit in a GTP-dependent manner and the cells depleted of Der accumulate the structurally unstable 50S subunit, which dissociates into an aberrant subunit at a lower Mg 2+ concentration. As Der is an essential and ubiquitous protein in bacteria, it may prove to be an ideal cellular target against which new antibiotics can be developed. In the present study, we describe our attempts to identify novel antibiotics specifically targeting Der GTPase. We performed the structure-based design of Der inhibitors using the X-ray crystal structure of Thermotoga maritima Der (TmDer). Virtual screening of commercially available chemical library retrieved 257 small molecules that potentially inhibit Der GTPase activity. These 257 chemicals were tested for their in vitro effects on TmDer GTPase and in vivo antibacterial activities. We identified three structurally diverse compounds, SBI-34462, -34566 and -34612, that are both biologically active against bacterial cells and putative enzymatic inhibitors of Der GTPase homologs. We also presented the possible interactions of each compound with the Der GTP-binding site to understand the mechanism of inhibition. Therefore, our lead compounds inhibiting Der GTPase provide scaffolds for the development of novel antibiotics against antibiotic-resistant pathogenic bacteria.

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“…Zantrin Z3 (37 in Figure 6), a quinazoline derivative, was reported as a GTPase inhibitor of FtsZ with an IC 50 of 24 μM by Margalit and co-workers through a high throughput screening assay [111]. This compound was reported to perturb Z-ring assembly in E. coli cells and cause lethality to a variety of bacteria in broth cultures [111].…”

Section: Quinazoline Derivativesmentioning

confidence: 99%

“…This compound was reported to perturb Z-ring assembly in E. coli cells and cause lethality to a variety of bacteria in broth cultures [111]. To develop reliable and valuable new FtsZ inhibitors, Shaw and co-workers preformed a broad biochemical crosscomparison of many known FtsZ inhibitors for the selection of reliable molecular scaffolds for further structural advancement.…”

Section: Quinazoline Derivativesmentioning

confidence: 99%