The structural assembly switch of cell division protein FtsZ probed with fluorescent allosteric inhibitors

Artola, Marta; Ruiz-Avila, Laura B.; Ramírez-Aportela, Erney; Martínez, R. Fernando; Araújo‐Bazán, Lidia; Vázquez-Villa, Henar; Martín‐Fontecha, Mar; Oliva, M.A.; Martín-Galiano, Antonio J.; Chacón, Pablo; López-Rodrı́guez, Marı́a L.; Andreu, José Manuel; Huecas, Sonia

doi:10.1039/c6sc03792e

Cited by 37 publications

(53 citation statements)

References 59 publications

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“…When we grew this strain in the presence of cadmium to induce mciZ expression we observed enlarged "ballooning" cells, a phenotype characteristic of blocking cell division in cocci (Pinho and Errington, 2003;Haydon et al, 2008). Additionally, using a fluorescent probe that we had developed to label FtsZ polymers (Artola et al, 2017), we could confirm the absence of Z-rings in these enlarged cells (Fig. 8B).…”

Section: Inhibition Of S Aureus Cell Division By Intracellular Exprementioning

confidence: 86%

“…BsFtsZ, BsFtsZCt (Huecas et al, 2017), SaFtsZ and EcFtsZ (Artola et al, 2017) were purified as described. His 6 -BsFtsZ and His 6 -BsFtsZΔCt were purified as BsFtsZ and BsFtsZΔCt, respectively, but without the His-tag cleavage step.…”

Section: Protein Purificationmentioning

confidence: 99%

“…FtsZ is a tubulin-like GTPase whose subunits assemble head-to-tail forming polar filaments (Nogales et al, 1998;Matsui et al, 2012) that laterally associate into bundles through the disordered C-terminal tails (Buske et al, 2015;Huecas et al, 2017;Sundararajan et al, 2018). GTP hydrolysis at the interface between monomers in filaments and a polymerization-driven structural switch enable FtsZ filament treadmilling dynamics (Artola et al, 2017;Wagstaff et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

“…FtsZ inhibition blocks cell division, inducing the formation of larger undivided cells that subsequently die. Many experimental small molecule FtsZ inhibitors have been reported (Schaffner-Barbero et al, 2012;den Blaauwen et al, 2014), including GTP-replacing inhibitors (Ruiz-Avila et al, 2013;Artola et al, 2015) and allosteric modulators of the structural assembly switch (Elsen et al, 2012;Artola et al, 2017). The difluorobenzamide derivative PC190723 (Haydon et al, 2008) and several of its analogs are currently the best characterized FtsZ-targeting antibacterial inhibitors, including biochemical, structural, cellular and resistance mechanisms (Andreu et al, 2010;Matsui et al, 2012;Tan et al, 2012;Stokes et al, 2013;Adams et al, 2016;Kaul et al, 2016;Fujita et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

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Synthetic developmental regulator MciZ targets FtsZ across Bacillus species and inhibits bacterial division

Araújo‐Bazán

Huecas

Valle

et al. 2019

Molecular Microbiology

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Summary Cell division in most bacteria is directed by FtsZ, a conserved tubulin‐like GTPase that assembles forming the cytokinetic Z‐ring and constitutes a target for the discovery of new antibiotics. The developmental regulator MciZ, a 40‐amino acid peptide endogenously produced during Bacillus subtilis sporulation, halts cytokinesis in the mother cell by inhibiting FtsZ. The crystal structure of a FtsZ:MciZ complex revealed that bound MciZ extends the C‐terminal β‐sheet of FtsZ blocking its assembly interface. Here we demonstrate that exogenously added MciZ specifically inhibits B. subtilis cell division, sporulation and germination, and provide insight into MciZ molecular recognition by FtsZ from different bacteria. MciZ and FtsZ form a complex with sub‐micromolar affinity, analyzed by analytical ultracentrifugation, laser biolayer interferometry and isothermal titration calorimetry. Synthetic MciZ analogs, carrying single amino acid substitutions impairing MciZ β‐strand formation or hydrogen bonding to FtsZ, show a gradual reduction in affinity that resembles their impaired activity in bacteria. Gene sequences encoding MciZ spread across genus Bacillus and synthetic MciZ slows down cell division in Bacillus species, including pathogenic Bacillus cereus and Bacillus anthracis. Moreover, B. subtilis MciZ is recognized by the homologous FtsZ from Staphylococcus aureus and inhibits division when it is expressed into S. aureus cells.

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Section: Inhibition Of S Aureus Cell Division By Intracellular Exprementioning

confidence: 86%

Section: Protein Purificationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Synthetic developmental regulator MciZ targets FtsZ across Bacillus species and inhibits bacterial division

Araújo‐Bazán

Huecas

Valle

et al. 2019

Molecular Microbiology

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“…FtsZ is a tubulin-like GTPase that recruit cell division proteins for new cell wall formation [162,163]. Due to its pivotal role in cell division, it has been recognised as an important target for various antibacterial compounds or drugs including natural products.…”

Section: Cell Division Protein Ftszmentioning

confidence: 99%

Non-microbial Natural Products That Inhibit Drug-Resistant Staphylococcus aureus

Chew¹,

Peh²,

Yeang³

2019

Staphylococcus Aureus

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Drug resistance developed in human pathogenic bacteria is emerging and has become a global problem. Methicillin-resistant Staphylococcus aureus (MRSA) spreading in both hospital and community areas has posed a great impact to global public health. Current antibiotics used against these resistant strains are no longer efficacious and the search for new alternative is in urgent need. In the past decades, natural products have demonstrated multiple biological activities in biomedical areas including their antibacterial actions against various drug-resistant bacteria. More promisingly, some natural products could reverse the resistance of bacteria to the antibiotics, making the target bacteria susceptible to these drugs again. Numerous natural products have also exhibited potent synergism against the drug-resistant bacteria when used in combination with various types of antibiotics. Recently, several antibacterials derived from microbes have been developed and approved by Food and Drug Administration (FDA) for clinical use. In this chapter, we discuss the potential use of non-microbial natural products in controlling Staphylococcus aureus (S. aureus)'s growth, and the underlying challenges in developing the natural products into clinical applications.

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Evidence of conformational switch in Streptococcus pneumoniae FtsZ during polymerization

et al. 2021

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FtsZ, the master coordinator of bacterial cell division, assembles into filaments in the presence of nucleotide. FtsZ from Streptococcus pneumoniae bears two tryptophan residues (W294 and W378) in its amino acid sequence. The tryptophan fluorescence of FtsZ increases during the assembly of FtsZ. We hypothesized that this increase in the fluorescence intensity was due to the change in the environment of one or both tryptophan residues. To examine this, we constructed two mutants (W294F and W378F) of FtsZ by individually replacing tryptophan with phenylalanine. The mutants displayed similar secondary structures, GTPase activity, and polymerization ability as the wild type FtsZ. During the polymerization, only one tryptophan (W294) showed an increase in its fluorescence intensity. Using time‐correlated single‐photon counting, the fluorescence lifetime of W294 was found to be significantly higher than W378, indicating that W294 was more buried in the structure than W378. The lifetime of W294 further increased during polymer formation, while that of W378 remained unchanged. Fluorescence quenching experiment suggested that the solvent exposure of W294 reduced during the polymerization of FtsZ. W294 is located near the T‐7 loop of the protein, a region important for the monomer‐monomer interaction during the formation of a protofilament. The results indicated that the region around W294 of S. pneumoniae FtsZ undergoes a conformational switch during polymerization as seen for FtsZ from other bacteria.

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The structural assembly switch of cell division protein FtsZ probed with fluorescent allosteric inhibitors

Abstract: The assembly switch of cell division protein FtsZ monitored with a fluorescent probe targeting the cleft between domains.

Cited by 37 publications

References 59 publications

Synthetic developmental regulator MciZ targets FtsZ across Bacillus species and inhibits bacterial division

Synthetic developmental regulator MciZ targets FtsZ across Bacillus species and inhibits bacterial division

Non-microbial Natural Products That Inhibit Drug-Resistant Staphylococcus aureus

Evidence of conformational switch in Streptococcus pneumoniae FtsZ during polymerization

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