RT Journal Article
T1 The structural assembly switch of cell division protein FtsZ probed with fluorescent allosteric inhibitors
A1 Artola Pérez de Azanza, Marta Elena
A1 Ruiz Ávila, Laura, Laura B.
A1 Ramírez Aportela, Erney
A1 Fernando Martínez, R.
A1 Araujo Bazán, Lidia
A1 Vázquez Villa, Henar
A1 Martín Fontecha, María del Mar
A1 Oliva Blanco, María Ángela
A1 Martín Galiano, Antonio Javier
A1 Chacón Montes, Pablo
A1 López Rodríguez, María Luz
A1 Andreu Rodríguez, José Manuel
A1 Huecas Gayo, Sonia
AB FtsZ is a widely conserved tubulin-like GTPase that directs bacterial cell division and a new target for antibiotic discovery. This protein assembly machine cooperatively polymerizes forming single-stranded filaments, by means of self-switching between inactive and actively associating monomer conformations. The structural switch mechanism was proposed to involve a movement of the C-terminal and N-terminal FtsZ domains, opening a cleft between them, allosterically coupled to the formation of a tight association interface between consecutive subunits along the filament. The effective antibacterial benzamide PC190723 binds into the open interdomain cleft and stabilizes FtsZ filaments, thus impairing correct formation of the FtsZ ring for cell division. We have designed fluorescent analogs of PC190723 to probe the FtsZ structural assembly switch. Among them, nitrobenzoxadiazole probes specifically bind to assembled FtsZ rather than to monomers. Probes with several spacer lengths between the fluorophore and benzamide moieties suggest a binding site extension along the interdomain cleft. These probes label FtsZ rings of live Bacillus subtilis and Staphylococcus aureus, without apparently modifying normal cell morphology and growth, but at high concentrations they induce impaired bacterial division phenotypes typical of benzamide antibacterials. During the FtsZ assembly-disassembly process, the fluorescence anisotropy of the probes changes upon binding and dissociating from FtsZ, thus reporting open and closed FtsZ interdomain clefts. Our results demonstrate the structural mechanism of the FtsZ assembly switch, and suggest that the probes bind into the open clefts in cellular FtsZ polymers preferably to unassembled FtsZ in the bacterial cytosol.
PB RSC
SN 2041-6520
YR 2017
FD 2017
LK https://hdl.handle.net/20.500.14352/18293
UL https://hdl.handle.net/20.500.14352/18293
LA eng
NO The article was received on 24 Aug 2016, accepted on 19 Oct 2016 and first published on 21 Oct 2016
NO Ministerio de Economía y Competitividad de España (MINECO)
NO CSIC
DS Docta Complutense
RD 11 jul 2024