Escherichia coli FtsA forms lipid-bound minirings that antagonize lateral interactions between FtsZ protofilaments

Krupka, Marcin; Rowlett, Veronica W.; Morado, Dustin R.; Vitrac, Heidi; Schoenemann, Kara M.; Li, Jun; Margolin, William

doi:10.1038/ncomms15957

Cited by 73 publications

(136 citation statements)

References 80 publications

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“…A–B). In addition, the presence or absence of ATP in the reactions did not have reproducible effects on the structures formed, as was observed previously (Szwedziak et al , ; Krupka et al , ). We then asked whether adding higher concentrations of proteins might drive their oligomerization into different structures, including more minirings.…”

Section: Resultssupporting

confidence: 80%

“…As FtsA* forms short arcs when bound to the lipid monolayer instead of minirings, we showed that this structural change removes the constraints on miniring‐bound FtsZ protofilaments to laterally interact, allowing FtsZ protofilaments to bundle (Krupka et al , ). Considering that three of the FtsA*‐like mutants also bypass ZipA and form alternative structures on lipid monolayers, we predicted that the mutant proteins would also allow FtsZ filaments to bundle.…”

Section: Resultsmentioning

confidence: 90%

“…Indeed, recent evidence suggests that the oligomerization state of FtsA is important for regulating FtsZ protofilament bundling during early stages of cell division (Krupka et al , ) as well as progression of cell division. One model for proto‐ring function predicts that the arrival of ZipA to the Z‐ring disrupts FtsA oligomers (Pichoff et al , ).…”

Section: Introductionmentioning

confidence: 99%

“…Using a lipid monolayer system, we recently showed that FtsA assembles into lipid‐bound 12‐mer minirings that bind FtsZ protofilaments and keep them separated from one another (Krupka et al , ). This anti‐bundling activity of FtsA in vitro is consistent with in vivo phenotypes.…”

Section: Introductionmentioning

confidence: 99%

“…In support of FtsA* as a stimulator of FtsZ protofilament bundling, when FtsZ* is coassembled on lipids with FtsA*, it assembles into large sheets of multiple protofilaments, strongly suggesting a synergistic bundling effect. These data led to a model in which decreased interaction between FtsA* subunits within FtsA oligomers results in enhanced FtsZ filament bundling (Krupka et al , ). To test the model, it would be valuable to know whether the properties we observed for FtsA* could be generalized to other FtsA*‐like mutants.…”

Section: Introductionmentioning

confidence: 99%

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Gain‐of‐function variants of FtsA form diverse oligomeric structures on lipids and enhance FtsZ protofilament bundling

Schoenemann¹,

Krupka²,

Rowlett³

et al. 2018

Molecular Microbiology

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Escherichia coli requires FtsZ, FtsA and ZipA proteins for early stages of cell division, the latter two tethering FtsZ polymers to the cytoplasmic membrane. Hypermorphic mutants of FtsA such as FtsA* (R286W) map to the FtsA self-interaction interface and can bypass the need for ZipA. Purified FtsA forms closed minirings on lipid monolayers that antagonize bundling of FtsZ protofilaments, whereas FtsA* forms smaller oligomeric arcs that enable bundling. Here, we examined three additional FtsA*-like mutant proteins for their ability to form oligomers on lipid monolayers and bundle FtsZ. Surprisingly, all three formed distinct structures ranging from mostly arcs (T249M), a mixture of minirings, arcs and straight filaments (Y139D) or short straight double filaments (G50E). All three could form filament sheets at higher concentrations with added ATP. Despite forming these diverse structures, all three mutant proteins acted like FtsA* to enable FtsZ protofilament bundling on lipid monolayers. Synthesis of the FtsA*-like proteins in vivo suppressed the toxic effects of a bundling-defective FtsZ, exacerbated effects of a hyper-bundled FtsZ, and rescued some thermosensitive cell division alleles. Together, the data suggest that conversion of FtsA minirings into any type of non-miniring oligomer can promote progression of cytokinesis through FtsZ bundling and other mechanisms.

show abstract

Section: Resultssupporting

confidence: 80%

Section: Resultsmentioning

confidence: 90%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Gain‐of‐function variants of FtsA form diverse oligomeric structures on lipids and enhance FtsZ protofilament bundling

Schoenemann¹,

Krupka²,

Rowlett³

et al. 2018

Molecular Microbiology

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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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Probing Membrane-Associated Cytoskeletal Oligomers of the Bacterial Divisome by Electron Microscopy and Tomography

Hu,

Margolin

2023

Methods in Molecular Biology

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Escherichia coli FtsA forms lipid-bound minirings that antagonize lateral interactions between FtsZ protofilaments

Cited by 73 publications

References 80 publications

Gain‐of‐function variants of FtsA form diverse oligomeric structures on lipids and enhance FtsZ protofilament bundling

Gain‐of‐function variants of FtsA form diverse oligomeric structures on lipids and enhance FtsZ protofilament bundling

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

Probing Membrane-Associated Cytoskeletal Oligomers of the Bacterial Divisome by Electron Microscopy and Tomography

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