title: Assembly of archaeal cell division protein FtsZ and a GTPase-inactive mutant into double-stranded filaments.
abstract: We have studied the assembly and GTPase of purified FtsZ from the hyperthermophilic archaeon Methanococcus jannaschii, a structural homolog of eukaryotic tubulin, employing wild-type FtsZ, FtsZ-His6 (histidine-tagged FtsZ), and the new mutants FtsZ-W319Y and FtsZ-W319Y-His6, with light scattering, nucleotide analyses, electron microscopy, and image processing methods. This has revealed novel properties of FtsZ. The GTPase of archaeal FtsZ polymers is suppressed in Na+-containing buffer, generating stabilized structures that require GDP addition for disassembly. FtsZ assembly is polymorphic. Archaeal FtsZ(wt) assembles into associated and isolated filaments made of two parallel protofilaments with a 43 A longitudinal spacing between monomers, and this structure is also observed in bacterial FtsZ from Escherichia coli. The His6 extension facilitates the artificial formation of helical tubes and sheets. FtsZ-W319Y-His6 is an inactivated GTPase whose assembly remains regulated by GTP and Mg2+. It forms two-dimensional crystals made of symmetrical pairs of tubulin-like protofilaments, which associate in an antiparallel array (similarly to the known Ca2+-induced sheets of FtsZ-His6). In contrast to the lateral interactions of microtubule protofilaments, we propose that the primary assembly product of FtsZ is the double-stranded filament, one or several of which might form the dynamic Z ring during prokaryotic cell division.
authors.affiliation: Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Cientificas, Madrid, Spain.
authors.forename: María A
authors.initials: MA
authors.lastname: Oliva
authors.forename: Sonia
authors.initials: S
authors.lastname: Huecas
authors.forename: Juan M
authors.initials: JM
authors.lastname: Palacios
authors.forename: Jaime
authors.initials: J
authors.lastname: Martín-Benito
authors.forename: José M
authors.initials: JM
authors.lastname: Valpuesta
authors.forename: José M
authors.initials: JM
authors.lastname: Andreu
mesh_terms: D001105:Archaea / Q000378:metabolism*; D019843:Archaeal Proteins / Q000737:chemistry* / Q000378:metabolism*; D002455:Cell Division; D004926:Escherichia coli / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000737:chemistry / Q000235:genetics*; D006160:Guanosine Triphosphate / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D008027:Light; D008274:Magnesium / Q000494:pharmacology; D017017:Methanococcus / Q000201:enzymology; D008854:Microscopy, Electron; D008954:Models, Biological; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D011188:Potassium / Q000494:pharmacology; D012542:Scattering, Radiation; D012964:Sodium / Q000494:pharmacology; D013696:Temperature; D013997:Time Factors
chemical_list: D019843:Archaeal Proteins; C119586:FtsZ protein, Methanococcus jannaschii; D006160:Guanosine Triphosphate; D012964:Sodium; D020558:GTP Phosphohydrolases; D008274:Magnesium; D011188:Potassium