Treadmilling analysis reveals new insights into dynamic FtsZ ring architecture

Ramirez-Diaz, Diego A.; García‐Soriano, Daniela A.; Raso, Ana; Mücksch, Jonas; Feingold, Mario; Rivas, Germán; Schwille, Petra

doi:10.1371/journal.pbio.2004845

Cited by 98 publications

(150 citation statements)

References 38 publications

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“…[71,72] It is also a cytosolic protein and needs either FtsA or ZipA in order to attach to the membrane in vivo. [76] These results could be used in the field of bottom-up synthetic biology to design a minimal divisome consisting of only two proteins: ZipA acting as a membrane anchor to transmit the motor force generated by FtsZ polymerization as has been shown by vesicle shrinkage and membrane invagination in vitro. [57] ZipA is integrated into the membrane via an N-terminal domain and also contains a C-terminal FtsZ-binding domain, both connected via a flexible linker.…”

Section: The Prokaryotic Divisomementioning

confidence: 91%

“…One big player in this context and one of the best studied prokaryotic division proteins to date is FtsZ, which is required for septum formation but does not itself perform the septation process. [76] FtsZ shows outstanding treadmilling dynamics in vivo as well as in vitro in circular polymer structures with a diameter of ≈1 µm. [71,72] It is also a cytosolic protein and needs either FtsA or ZipA in order to attach to the membrane in vivo.…”

Section: The Prokaryotic Divisomementioning

confidence: 99%

“…It is highly conserved in the bacterial domain and the ancestor of eukaryotic tubulin. [76] In combination with recent studies in E. coli [77] and B. subtilis [78] on the correlation between treadmilling dynamics and peptidoglycan synthesis, a new focus on the FtsZ structure and its dynamics can now be formulated, as well as provide insight into the mechanical aspects of bacterial cell division. FtsA is part of the actin-family and contains an amphipathic helix that is believed to mediate its binding to the membrane.…”

Section: The Prokaryotic Divisomementioning

confidence: 99%

“…[57] ZipA is integrated into the membrane via an N-terminal domain and also contains a C-terminal FtsZ-binding domain, both connected via a flexible linker. [76,[79][80][81] [76] FtsZ shows outstanding treadmilling dynamics in vivo as well as in vitro in circular polymer structures with a diameter of ≈1 µm.…”

Section: The Prokaryotic Divisomementioning

confidence: 99%

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Functional Modules of Minimal Cell Division for Synthetic Biology

et al. 2019

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Section: The Prokaryotic Divisomementioning

confidence: 91%

Section: The Prokaryotic Divisomementioning

confidence: 99%

Section: The Prokaryotic Divisomementioning

confidence: 99%

Section: The Prokaryotic Divisomementioning

confidence: 99%

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Functional Modules of Minimal Cell Division for Synthetic Biology

et al. 2019

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“…FtsZ, a homolog of the eukaryotic cytoskeletal protein, tubulin [5,6], is the chief constituent of the machinery that coordinates the partitioning of the bacterial cells. FtsZ also displays treadmilling behavior, which emphasizes the importance of the GTPase activity for the formation and functioning of the Zring in bacteria [12][13][14]. The Zring formation is orchestrated by coordinated actions of several positive and negative regulators of FtsZ assembly [8][9][10][11].…”

Section: Introductionmentioning

confidence: 99%

Regulation of Streptococcus pneumoniae FtsZ assembly by divalent cations: paradoxical effects of Ca²⁺ on the nucleation and bundling of FtsZ polymers

et al. 2019

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The assembly and disassembly of the FtsZ ring drives the division of bacteria cells, including Streptococcus pneumoniae, which causes pneumonia and meningitis. In contrast to FtsZ from other bacterial species, Streptococcus pneumoniae (Spn) FtsZ contains two tryptophan residues. Here, we demonstrate that the assembly and disassembly of Streptococcus pneumoniae FtsZ (SpnFtsZ) monomers can be monitored by the intrinsic tryptophan fluorescence of FtsZ. We found that the assembly of SpnFtsZ is closely associated with its GTPase activity. Guanosine 5′‐[β,γ‐imido]triphosphate, a nonhydrolyzable analog of GTP, stabilized the FtsZ filaments without inducing their bundling. Using intrinsic tryptophan fluorescence, light scattering, and electron microscopy, we could differentiate the effects of divalent calcium and magnesium on the assembly of FtsZ. Though Mg2+ increased the stability of the FtsZ filaments, it could not prevent the disassembly of the filaments under conditions where GTP was limiting. Thus, our results indicate that Mg2+ primarily enhances the longitudinal assembly of FtsZ. Low concentrations of Ca2+ strongly promoted the bundling of FtsZ filaments and inhibited the disassembly of the filaments, suggesting that low concentrations of Ca2+ enhance the lateral interactions between the FtsZ filaments. Interestingly, Ca2+ delayed the nucleation process of FtsZ assembly, indicating that Ca2+ exerts paradoxical effects on the assembly of FtsZ. However, higher concentrations of Ca2+ did not enhance the bundling of FtsZ filaments. In addition, Ca2+ altered the secondary structure of FtsZ and increased the fluorescence of the FtsZ‐1‐anilinonaphthalene‐8‐sulfonic acid complex, indicating that Ca2+ induces conformational changes in FtsZ. The study provides an interesting insight into the assembly of SpnFtsZ and its regulation by divalent cations.

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Gene‐Directed FtsZ Ring Assembly Generates Constricted Liposomes with Stable Membrane Necks

Godino

Danelon

2023

Advanced Biology

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vesicles. With a limited number of division factors compared to eukaryotes, the Escherichia coli divisome has been at the center of numerous efforts attempting to split liposomes. [5][6][7][8][9][10][11][12] In vivo, a contractile ring composed of FtsZ, FtsA, and ZipA is formed at midcell. [13,14] FtsZ is a bacterial homologue of tubulin [15][16][17] that polymerizes into self-interacting filaments when bound to GTP. [18][19][20][21] FtsZ lacks a membrane targeting domain, and it is anchored to the cytoplasmic bilayer by FtsA and ZipA.In vitro reconstruction of FtsZ polymers, [15,22] FtsA and ZipA [23] in model membranes has contributed to elucidate the self-organization and dynamics of the E. coli division proteins. In 2008, Osawa and colleagues suggested that Z-rings could generate constriction forces within tubular liposomes in the absence of motor proteins. [5] In this work, FtsZ-mts (a chimeric FtsZ with a membrane targeting sequence fused to the C terminus) caused membrane invagination, though full constriction was not achieved. [5] The authors proposed that the conformational change of FtsZ from straight to curved filaments acts as the driving force. In a follow-up study, the same group claimed that the Z-ring stimulates membrane constriction in giant liposomes to the point of septation. [7] The use of FtsA*, a gain-of-function mutant of FtsA, [24] was required to achieve complete scission, as such division events were not observed with FtsZ-mts. Szwedziak and colleagues showed that purified FtsZ and FtsA from Thermotoga maritima form continuous filament rings encircling the membrane on the inside of small unilamellar vesicles, generating constriction sites but evidences of membrane abscission were not presented. [9] The authors proposed a scenario, where the sliding and shortening of overlapping FtsZ filaments constitute the main force generator. [9] Interestingly, Ramirez Diaz and colleagues reported that FtsZ filaments do not have a single curvature, [25] and that their intrinsic helical structure drives liposome deformation via torsional stress, a mechanism that is reliant on GTP hydrolysis. [12] With the aim to recapitulate a functional Z-ring from gene-encoded proteins in liposomes, our group has demonstrated that ring-like cytoskeletal structures led to membrane constriction into narrow necks connecting budding vesicles, [11] a phenotype that differs from that observed in previous studies by the more pronounced pinching of the liposome membrane. Although no division events could unambiguously be detected, our experimental design was not directed toward addressing Mimicking bacterial cell division in well-defined cell-free systems has the potential to elucidate the minimal set of proteins required for cytoskeletal formation, membrane constriction, and final abscission. Membrane-anchored FtsZ polymers are often regarded as a sufficient system to realize this chain of events. By using purified FtsZ and its membrane-binding protein FtsA or the gain-of-function mutant FtsA* expressed in PURE (Protein syn...

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Treadmilling analysis reveals new insights into dynamic FtsZ ring architecture

Cited by 98 publications

References 38 publications

Functional Modules of Minimal Cell Division for Synthetic Biology

Functional Modules of Minimal Cell Division for Synthetic Biology

Regulation of Streptococcus pneumoniae FtsZ assembly by divalent cations: paradoxical effects of Ca²⁺ on the nucleation and bundling of FtsZ polymers

Gene‐Directed FtsZ Ring Assembly Generates Constricted Liposomes with Stable Membrane Necks

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Treadmilling analysis reveals new insights into dynamic FtsZ ring architecture

Cited by 98 publications

References 38 publications

Functional Modules of Minimal Cell Division for Synthetic Biology

Functional Modules of Minimal Cell Division for Synthetic Biology

Regulation of Streptococcus pneumoniae FtsZ assembly by divalent cations: paradoxical effects of Ca2+ on the nucleation and bundling of FtsZ polymers

Gene‐Directed FtsZ Ring Assembly Generates Constricted Liposomes with Stable Membrane Necks

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Regulation of Streptococcus pneumoniae FtsZ assembly by divalent cations: paradoxical effects of Ca²⁺ on the nucleation and bundling of FtsZ polymers