scholarly journals Cell-free biogenesis of bacterial division proto-rings that can constrict liposomes

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Elisa Godino ◽  
Jonás Noguera López ◽  
Ilias Zarguit ◽  
Anne Doerr ◽  
Mercedes Jimenez ◽  
...  
Keyword(s):  
Cytoskeletal Proteins ◽  
Ftsz Ring ◽  
Division Site ◽  
Artificial Cell ◽  
Bacterial Division ◽  
Synthetic Cell ◽  
Free Gene ◽  
Bacterial Cell Cycle ◽  
Planar Membranes ◽  
Genetic Programme

AbstractA major challenge towards the realization of an autonomous synthetic cell resides in the encoding of a division machinery in a genetic programme. In the bacterial cell cycle, the assembly of cytoskeletal proteins into a ring defines the division site. At the onset of the formation of the Escherichia coli divisome, a proto-ring consisting of FtsZ and its membrane-recruiting proteins takes place. Here, we show that FtsA-FtsZ ring-like structures driven by cell-free gene expression can be reconstituted on planar membranes and inside liposome compartments. Such cytoskeletal structures are found to constrict the liposome, generating elongated membrane necks and budding vesicles. Additional expression of the FtsZ cross-linker protein ZapA yields more rigid FtsZ bundles that attach to the membrane but fail to produce budding spots or necks in liposomes. These results demonstrate that gene-directed protein synthesis and assembly of membrane-constricting FtsZ-rings can be combined in a liposome-based artificial cell.

scholarly journals Cell-free biogenesis of bacterial division proto-rings that can constrict liposomes

2020 ◽  
Author(s):  
Elisa Godino ◽  
Jonás Noguera López ◽  
Ilias Zarguit ◽  
Anne Doerr ◽  
Mercedes Jimenez ◽  
...  
Keyword(s):  
Cytoskeletal Proteins ◽  
Division Site ◽  
Artificial Cell ◽  
Bacterial Division ◽  
Synthetic Cell ◽  
Free Gene ◽  
Bacterial Cell Cycle ◽  
Cytoskeletal Elements ◽  
Planar Membranes ◽  
Genetic Programme

ABSTRACTA major challenge towards the realization of an autonomous synthetic cell resides in the encoding of a division machinery in a genetic programme. A key event in the bacterial cell cycle is the assembly of cytoskeletal proteins into a ring that defines the division site. At the onset of the formation of the Escherichia coli divisome, a proto-ring consisting of FtsZ and its membrane recruiting proteins takes place. Here, we show that FtsA-FtsZ ring-like structures driven by cell-free gene expression can be reconstituted on planar membranes and inside liposome compartments. Such cytoskeletal structures are found to constrict the membrane and generate budding vesicles, a phenotype that has not been reported before. Additional expression of the FtsZ cross-linker protein ZapA yields more rigid FtsZ bundles that attach to the membrane but fail to produce budding spots or necks in liposomes. These results provide new insights on the self-organization of basic cytoskeletal elements involved in bacterial division. Moreover, they demonstrate that gene-directed protein synthesis and assembly of membrane-constricting FtsZ-rings can be combined in a liposome-based artificial cell.

scholarly journals Effects of Perturbing Nucleoid Structure on Nucleoid Occlusion-Mediated Toporegulation of FtsZ Ring Assembly

2004 ◽  
Vol 186 (12) ◽  
pp. 3951-3959 ◽  
Author(s):  
Qin Sun ◽  
William Margolin
Keyword(s):  
Chromosome Segregation ◽  
Potential Role ◽  
Membrane Insertion ◽  
Translation Inhibition ◽  
Ftsz Ring ◽  
Division Site ◽  
Ring Assembly ◽  
Z Ring ◽  
Nucleoid Structure

ABSTRACT In Escherichia coli, assembly of the FtsZ ring (Z ring) at the cell division site is negatively regulated by the nucleoid in a phenomenon called nucleoid occlusion (NO). Previous studies have indicated that chromosome packing plays a role in NO, as mukB mutants grown in rich medium often exhibit FtsZ rings on top of diffuse, unsegregated nucleoids. To address the potential role of overall nucleoid structure on NO, we investigated the effects of disrupting chromosome structure on Z-ring positioning. We found that NO was mostly normal in cells with inactivated DNA gyrase or in mukB-null mutants lacking topA, although some suppression of NO was evident in the latter case. Previous reports suggesting that transcription, translation, and membrane insertion of proteins (“transertion”) influence nucleoid structure prompted us to investigate whether disruption of these activities had effects on NO. Blocking transcription caused nucleoids to become diffuse, and FtsZ relocalized to multiple bands on top of these nucleoids, biased towards midcell. This suggested that these diffuse nucleoids were defective in NO. Blocking translation with chloramphenicol caused characteristic nucleoid compaction, but FtsZ rarely assembled on top of these centrally positioned nucleoids. This suggested that NO remained active upon translation inhibition. Blocking protein secretion by thermoinduction of a secA(Ts) strain caused a chromosome segregation defect similar to that in parC mutants, and NO was active. Although indirect effects are certainly possible with these experiments, the above data suggest that optimum NO activity may require specific organization and structure of the nucleoid.

scholarly journals Assembly of bacterial cell division protein FtsZ into dynamic biomolecular condensates

2020 ◽  
Author(s):  
Miguel Ángel Robles-Ramos ◽  
Silvia Zorrilla ◽  
Carlos Alfonso ◽  
William Margolin ◽  
Germán Rivas ◽  
...  
Keyword(s):  
Cell Division ◽  
Bacterial Cell ◽  
Bound State ◽  
Bacterial Cell Division ◽  
Structural Element ◽  
Physiological Factors ◽  
General Role ◽  
E Coli ◽  
Division Site ◽  
Synthetic Cell

Biomolecular condensation through phase separation may be a novel mechanism to regulate bacterial processes, including cell division. Previous work revealed FtsZ, a protein essential for cytokinesis in most bacteria, and the E. coli division site selection factor SlmA form FtsZ∙SlmA biomolecular condensates. The absence of condensates composed solely of FtsZ under the conditions used in that study suggested this mechanism was restricted to nucleoid occlusion or SlmA-containing bacteria. Here we report that FtsZ alone can demix into condensates in bulk and when encapsulated in synthetic cell-like systems. Condensate assembly depends on FtsZ being in the GDP-bound state and on crowding conditions that promote its oligomerization. FtsZ condensates are dynamic and gradually convert into FtsZ filaments upon GTP addition. Notably, FtsZ lacking its C-terminal disordered region, a structural element likely to favor biomolecular condensation, also forms condensates, albeit less efficiently. The inherent tendency of FtsZ to form condensates susceptible to modulation by physiological factors, including binding partners, suggests that such mechanisms may play a more general role in bacterial cell division than initially envisioned.

Synthetic organelles

2019 ◽  
Vol 3 (5) ◽  
pp. 587-595 ◽  
Author(s):  
Friedrich C. Simmel
Keyword(s):  
Short Review ◽  
Review Article ◽  
Chemical Processes ◽  
Enzyme Reactions ◽  
Chemical Gradients ◽  
Cell Structures ◽  
Synthetic Cell ◽  
Recent Developments ◽  
Free Gene ◽  
Specific Synthesis

One approach towards the creation of bottom-up synthetic biological systems of higher complexity relies on the subcompartmentalization of synthetic cell structures using artificially generated organelles — roughly mimicking the architecture of eukaryotic cells. Organelles create dedicated chemical environments for specific synthesis tasks — they separate incompatible processes from each other and help to create or maintain chemical gradients that drive other chemical processes. Artificial organelles have been used to compartmentalize enzyme reactions, to generate chemical fuels via photosynthesis and oxidative phosphorylation, and they have been utilized to spatially organize cell-free gene expression reactions. In this short review article, we provide an overview of recent developments in this field, which involve a wide variety of compartmentalization strategies ranging from lipid and polymer membrane systems to membraneless compartmentalization via coacervation.

scholarly journals MapZ forms a stable ring structure that acts as a nano-track for FtsZ treadmilling inStreptococcus mutans

2017 ◽  
Author(s):  
Yongliang Li ◽  
Shipeng Shao ◽  
Xiao Xu ◽  
Xiaodong Su ◽  
Yujie Sun ◽  
...  
Keyword(s):  
Single Molecule ◽  
Super Resolution ◽  
Ring Structure ◽  
Antibacterial Drug ◽  
Ftsz Ring ◽  
Division Site ◽  
Binary Division ◽  
Z Ring ◽  
Vital Component ◽  
Stable Ring

AbstractBacterial binary division requires the accurate placement of the division machinery. FtsZ, the vital component of the division machinery, can assemble into filaments and self-organize into a ring structure (Z-ring) at the proper site for cell division. Thus, understanding how bacteria control the spatiotemporal formation of the FtsZ ring is crucial for small molecule and nanoparticle antibacterial drug discovery. MapZ, a recently identified FtsZ regulator inStreptococcaceae,has been found to localize at the mid-cell and position the FtsZ ring. However, the mechanism is still unclear. Here, by using total internal reflection fluorescence microscopy, super-resolution imaging, and single molecule tracking, we investigated the mechanism by which MapZ regulates the FtsZ ring position. The results show that FtsZ exhibites dynamic treadmilling motion in S.mutans.Importantly, depletion of MapZ leads to an unconstrained movement of treadmilling FtsZ filaments and a shorter lifetime of the constricting FtsZ ring. Furthermore, by revealing that MapZ forms an immobile ring-like nanostructure at the division site, our study suggests that MapZ forms a stable ring that acts as a nanotrack to guide and restrict treadmilling FtsZ filaments in S.mutans, representing a novel way in which bacteria control the division.

scholarly journals PomX, a ParA/MinD ATPase activating protein, is a triple regulator of cell division in Myxococcus xanthus

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Dominik Schumacher ◽  
Andrea Harms ◽  
Silke Bergeler ◽  
Erwin Frey ◽  
Lotte Sogaard-Andersen
Keyword(s):  
Cell Division ◽  
Myxococcus Xanthus ◽  
Dependent Manner ◽  
Ftsz Ring ◽  
Work Support ◽  
Terminal Domain ◽  
Division Site ◽  
The Social ◽  
Underlying Mechanisms ◽  
Positively Charged

Cell division site positioning is precisely regulated but the underlying mechanisms are incompletely understood. In the social bacterium Myxococcus xanthus, the ~15 MDa tripartite PomX/Y/Z complex associates with and translocates across the nucleoid in a PomZ ATPase-dependent manner to directly position and stimulate formation of the cytokinetic FtsZ-ring at midcell, and then undergoes fission during division. Here, we demonstrate that PomX consists of two functionally distinct domains and has three functions. The N-terminal domain stimulates ATPase activity of the ParA/MinD ATPase PomZ. The C-terminal domain interacts with PomY and forms polymers, which serve as a scaffold for PomX/Y/Z complex formation. Moreover, the PomX/PomZ interaction is important for fission of the PomX/Y/Z complex. These observations together with previous work support that the architecturally diverse ATPase activating proteins of ParA/MinD ATPases are highly modular and use the same mechanism to activate their cognate ATPase via a short positively charged N-terminal extension.

scholarly journals The Assembly of the FtsZ Ring at the Mid-Chloroplast Division Site Depends on a Balance Between the Activities of AtMinE1 and ARC11/AtMinD1

2008 ◽  
Vol 49 (3) ◽  
pp. 345-361 ◽  
Author(s):  
Makoto T. Fujiwara ◽  
Haruki Hashimoto ◽  
Yusuke Kazama ◽  
Tomoko Abe ◽  
Shigeo Yoshida ◽  
...  
Keyword(s):  
Chloroplast Division ◽  
Ftsz Ring ◽  
Division Site

scholarly journals Growth Rate-Dependent Regulation of Medial FtsZ Ring Formation

2003 ◽  
Vol 185 (9) ◽  
pp. 2826-2834 ◽  
Author(s):  
Richard B. Weart ◽  
Petra Anne Levin
Keyword(s):  
Cell Cycle ◽  
Growth Rate ◽  
Transcriptional Control ◽  
Doubling Time ◽  
Ring Formation ◽  
Intracellular Concentration ◽  
Dependent Manner ◽  
Ftsz Ring ◽  
Division Site ◽  
Rate Dependent

ABSTRACT FtsZ is an essential cell division protein conserved throughout the bacteria and archaea. In response to an unknown cell cycle signal, FtsZ polymerizes into a ring that establishes the future division site. We conducted a series of experiments examining the link between growth rate, medial FtsZ ring formation, and the intracellular concentration of FtsZ in the gram-positive bacterium Bacillus subtilis. We found that, although the frequency of cells with FtsZ rings varies as much as threefold in a growth rate-dependent manner, the average intracellular concentration of FtsZ remains constant irrespective of doubling time. Additionally, expressing ftsZ solely from a constitutive promoter, thereby eliminating normal transcriptional control, did not alter the growth rate regulation of medial FtsZ ring formation. Finally, our data indicate that overexpressing FtsZ does not dramatically increase the frequency of cells with medial FtsZ rings, suggesting that the mechanisms governing ring formation are refractile to increases in FtsZ concentration. These results support a model in which the timing of FtsZ assembly is governed primarily through cell cycle-dependent changes in FtsZ polymerization kinetics and not simply via oscillations in the intracellular concentration of FtsZ. Importantly, this model can be extended to the gram-negative bacterium Escherichia coli. Our data show that, like those in B. subtilis, average FtsZ levels in E. coli are constant irrespective of doubling time.

scholarly journals ARC3 Activation by PARC6 Promotes FtsZ-Ring Remodeling at the Chloroplast Division Site

2019 ◽  
Vol 31 (4) ◽  
pp. 862-885
Author(s):  
Cheng Chen ◽  
Lingyan Cao ◽  
Yue Yang ◽  
Katie J. Porter ◽  
Katherine W. Osteryoung
Keyword(s):  
Chloroplast Division ◽  
Ftsz Ring ◽  
Division Site

scholarly journals The Division Inhibitor EzrA Contains a Seven-Residue Patch Required for Maintaining the Dynamic Nature of the Medial FtsZ Ring

2007 ◽  
Vol 189 (24) ◽  
pp. 9001-9010 ◽  
Author(s):  
Daniel P. Haeusser ◽  
Anna Cristina Garza ◽  
Amy Z. Buscher ◽  
Petra Anne Levin
Keyword(s):  
De Novo ◽  
Dependent Manner ◽  
Dynamic Nature ◽  
Ftsz Ring ◽  
Division Site ◽  
C Terminus ◽  
Ftsz Assembly ◽  
Mutant Cells ◽  
Precise Role

ABSTRACT The essential cytoskeletal protein FtsZ assembles into a ring-like structure at the nascent division site and serves as a scaffold for the assembly of the prokaryotic division machinery. We previously characterized EzrA as an inhibitor of FtsZ assembly in Bacillus subtilis. EzrA interacts directly with FtsZ to prevent aberrant FtsZ assembly and cytokinesis at cell poles. EzrA also concentrates at the cytokinetic ring in an FtsZ-dependent manner, although its precise role at this position is not known. Here, we identified a conserved patch of amino acids in the EzrA C terminus that is essential for localization to the FtsZ ring. Mutations in this patch (designated the “QNR patch”) abolish EzrA localization to midcell but do not significantly affect EzrA's ability to inhibit FtsZ assembly at cell poles. ezrA QNR patch mutant cells exhibit stabilized FtsZ assembly at midcell and are significantly longer than wild-type cells, despite lacking extra FtsZ rings. These results indicate that EzrA has two distinct activities in vivo: (i) preventing aberrant FtsZ ring formation at cell poles through inhibition of de novo FtsZ assembly and (ii) maintaining proper FtsZ assembly dynamics within the medial FtsZ ring, thereby rendering it sensitive to the factors responsible for coordinating cell growth and cell division.

Sign in / Sign up

Export Citation Format

Share Document