FtsZ polymers bound to lipid bilayers through ZipA form dynamic two dimensional networks

Mateos-Gil, Pablo; Márquez, Ileana; López-Navajas, Pilar; Jiménez, Mercedes; Vicente, Miguel; Mingorance, Jesús; Rivas, Germán; Vélez, Marisela

doi:10.1016/j.bbamem.2011.12.012

Cited by 41 publications

(57 citation statements)

References 43 publications

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“…This allowed us to attach the rest of the protein to a bilayer containing Ni-chelating lipids, thereby mimicking the permanent membrane attachment of full-length ZipA (ref. 42). Like in our experiments with FtsA, we first incubated the membrane with both proteins and then initiated FtsZ polymerization by adding GTP.…”

Section: Resultsmentioning

confidence: 99%

The bacterial cell division proteins FtsA and FtsZ self-organize into dynamic cytoskeletal patterns

2013

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Bacterial cytokinesis is commonly initiated by the Z-ring, a cytoskeletal structure assembling at the site of division. Its primary component is FtsZ, a tubulin superfamily GTPase, which is recruited to the membrane by the actin-related protein FtsA. Both proteins are required for the formation of the Z-ring, but if and how they influence each other's assembly dynamics is not known. Here, we reconstituted FtsA-dependent recruitment of FtsZ polymers to supported membranes, where both proteins self-organize into complex patterns, such as fast-moving filament bundles and chirally rotating rings. Using fluorescence microscopy and biochemical perturbations, we found that these large-scale rearrangements of FtsZ emerge from its polymerization dynamics and a dual, antagonistic role of FtsA: recruitment of FtsZ filaments to the membrane and a negative regulation on FtsZ organization. Our findings provide a model for the initial steps of bacterial cell division and illustrate how dynamic polymers can self-organize into large-scale structures.As in eukaryotic cells, proteins related to actin and tubulin provide the key structural components coordinating cellular functions in bacteria 1,2 . For example, cell division in most bacteria depends on the tubulin-related GTPase FtsZ and the widely conserved actin-related protein FtsA, which form an annular structure at the middle of the cell 3,4 . Purified FtsZ assembles into polar, straight or gently curved protofilaments in the presence of GTP [5][6][7][8] and lateral interactions between FtsZ protofilaments can lead to higher-ordered structures, like tubules, bundles, circles and sheets 5,6 . In most bacteria, FtsZ is recruited to the membrane by FtsA, which binds to the membrane via a C-terminal amphipathic helix 9,10 . Although binding of ATP is required for FtsA to interact with FtsZ, no ATPase activity of FtsA was found [9][10][11][12] . In Escherichia coli and other Gammaproteobacteria, FtsZ is also recruited to the membrane by the trans-membrane protein ZipA 13-15 and both membrane anchors are required for successful cell division. Although structurally not related, FtsA and ZipA bind to the same C-terminal peptide of FtsZ, which is connected to the rest of the protein via a flexible linker 16,17 . Most previous models for Z-ring formation mainly assumed both proteins to be passive membrane anchors for FtsZ [18][19][20] , an idea also followed in reconstitution studies, where the requirement for physiological membrane anchors was circumvented by supplying FtsZ with its own membrane targeting peptide [21][22][23] . However, in vitro experiments using an FtsA mutant suggested that the membrane anchor can change the * To whom correspondence should be addressed: martin_loose@hms.harvard.edu (M.L.). NIH Public Access RESULTS FtsZ and FtsA self-organize into a rapidly reorganizing filament networkEarlier attempts to reconstitute FtsA-FtsZ interaction were frustrated by wildtype FtsA being notoriously difficult to purify. Here, we circumvented this problem by using a S...

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Section: Resultsmentioning

confidence: 99%

The bacterial cell division proteins FtsA and FtsZ self-organize into dynamic cytoskeletal patterns

2013

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“…However, the function of ZipA was challenged by recent studies. First, ZipA induced FtsZ bundling was not significant at pH greater than 7 13 . Second, as referenced above, some FtsA mutants are able to bypass the need of ZipA.…”

Section: Introductionmentioning

confidence: 91%

ZipA and FtsA* stabilize FtsZ-GDP miniring structures

Chen

Huang

Osawa

et al. 2017

Sci Rep

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The cytokinetic division ring of Escherichia coli comprises filaments of FtsZ tethered to the membrane by FtsA and ZipA. Previous results suggested that ZipA is a Z-ring stabilizer, since in vitro experiments it is shown that ZipA enhanced FtsZ assembly and caused the filaments to bundles. However, this function of ZipA has been challenged by recent studies. First, ZipA-induced FtsZ bundling was not significant at pH greater than 7. Second, some FtsA mutants, such as FtsA* were able to bypass the need of ZipA. We reinvestigated the interaction of FtsZ with ZipA in vitro. We found that ZipA not only stabilized and bundled straight filaments of FtsZ-GTP, but also stabilized the highly curved filaments and miniring structures formed by FtsZ-GDP. FtsA* had a similar stabilization of FtsZ-GDP minirings. Our results suggest that ZipA and FtsA* may contribute to constriction by stabilizing this miniring conformation.

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“…Recent experiments describing the behavior of FtsZ filaments attached to supported lipid bilayers (29) indicate that the essential traits of the dynamic behavior of the FtsZ filaments observed on mica (15) are preserved on lipid surfaces, in spite of the formation of higher order aggregates. Accordingly, experimental evidence shows that the FtsZ ring in cells retains a dynamic behavior (30).…”

Section: Discussionmentioning

confidence: 99%

Depolymerization dynamics of individual filaments of bacterial cytoskeletal protein FtsZ

Mateos-Gil

Paez²,

Hörger³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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We report observation and analysis of the depolymerization filaments of the bacterial cytoskeletal protein FtsZ (filament temperature-sensitive Z) formed on a mica surface. At low concentration, proteins adsorbed on the surface polymerize forming curved filaments that close into rings that remain stable for some time before opening irreversibly and fully depolymerizing. The distribution of ring lifetimes (T ) as a function of length (N), shows that the rate of ring aperture correlates with filament length. If this ring lifetime is expressed as a bond survival time, (T b ≡ NT), this correlation is abolished, indicating that these rupture events occur randomly and independently at each monomer interface. After rings open irreversibly, depolymerization of the remaining filaments is fast, but can be slowed down and followed using a nonhydrolyzing GTP analogue. The histogram of depolymerization velocities of individual filaments has an asymmetric distribution that can be fit with a computer model that assumes two rupture rates, a slow one similar to the one observed for ring aperture, affecting monomers in the central part of the filaments, and a faster one affecting monomers closer to the open ends. From the quantitative analysis, we conclude that the depolymerization rate is affected both by nucleotide hydrolysis rate and by its exchange along the filament, that all monomer interfaces are equally competent for hydrolysis, although depolymerization is faster at the open ends than in central filament regions, and that all monomer-monomer interactions, regardless of the nucleotide present, can adopt a curved configuration.bacterial division | atomic force microscopy | computer simulations | GTP hydrolysis B acterial cytoskeletal protein FtsZ (filamenting temperaturesensitive Z) assembles into dynamic polymers on the inner side of the bacterial cytoplasmic membrane in the presence of GTP. Its role in cell division is twofold: recruiting the rest of the proteins involved in the assembly of the septal ring and contributing to generate the force needed for cell rupture. Recent in vitro experiments with artificially membrane-bound FtsZ reconstituted in model systems support the suggestion that FtsZ is capable of generating a contractile force without the need of any other proteins (1, 2). Several theoretical models trying to explain the molecular mechanism by which the GTP-dependent self-assembling polymerization process generates a contractile force have been recently proposed (3-12), but the debate is still open. Existing controversy over the relative contribution of filament curvature and lateral interactions in the constriction process is difficult to resolve until new experimental data determine how GTP hydrolysis is associated to monomer exchange within filaments.FtsZ is known to polymerize above a critical concentration in the presence of GTP (13). Bulk experiments in solution using a fluorescence signal to follow polymerization (14) have provided indirect evidence of monomer exchange, annealing, and depolymerizatio...

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FtsZ polymers bound to lipid bilayers through ZipA form dynamic two dimensional networks

Cited by 41 publications

References 43 publications

The bacterial cell division proteins FtsA and FtsZ self-organize into dynamic cytoskeletal patterns

The bacterial cell division proteins FtsA and FtsZ self-organize into dynamic cytoskeletal patterns

ZipA and FtsA* stabilize FtsZ-GDP miniring structures

Depolymerization dynamics of individual filaments of bacterial cytoskeletal protein FtsZ

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