Dynamics of bacterial cell division: Z ring condensation is essential for cytokinesis

Squyres, Georgia R.; Holmes, Matthew J; Barger, Sarah R.; Pennycook, Betheney R.; Ryan, Joël; Yan, Victoria Tianjing; Garner, Edward B.

doi:10.1101/2020.06.30.180737

Cited by 10 publications

(12 citation statements)

References 51 publications

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“…Consistent with this possibility, we observed that the average dwell time and percentage of the stationary FtsW-RFP population increased as the treadmilling speed of FtsZ decreased in GTPase mutant strains (Fig. 2f, Supplementary Table 4), in line with the observed increase in the dwell time of FtsZ monomers and FtsZ filament length in FtsZ GTPase mutants in a recent study (20). Interestingly, the average dwell time of stationary FtsW-RFP in wt cells (19.2±1.1s, Fig.…”

Section: Resultssupporting

confidence: 89%

“…The average dwell time of stationary FtsW-RFP molecules, however, continued to decrease to ~10s (Fig. 4e, blue squares), approaching the mean lifetime of FtsZ monomers in the Z-ring (8.1 sec)(20).…”

Section: Resultsmentioning

confidence: 91%

“…Interestingly, the average dwell time of stationary FtsW-RFP in wt cells (19.2±1.1s, Fig. 2f, Supplementary Table 4) is significantly longer than the lifetime of FtsZ monomers (8.1±0.5s) (20), indicating that other mechanism(s) may also be at play to maintain this stationary population of FtsW-RFP, as we will further describe below.…”

Section: Resultsmentioning

confidence: 92%

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FtsW exhibits distinct processive motions driven by either septal cell wall synthesis or FtsZ treadmilling in E. coli

Yang¹,

McQuillen

Lyu

et al. 2019

Preprint

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17 During bacterial cell division, synthesis of new septal peptidoglycan (sPG) is crucial for 18 successful cytokinesis and cell pole morphogenesis. FtsW, a SEDS (Shape, Elongation, Division and 19 Sporulation) family protein and an indispensable component of the cell division machinery in all 20 walled bacterial species, was recently identified in vitro as a new monofunctional peptidoglycan 21 glycosyltransferases (PGTase). FtsW and its cognate monofunctional transpeptidase (TPase) class b 22 penicillin binding protein (PBP3 or FtsI in E. coli) may constitute the essential, bifunctional sPG 23 synthase specific for new sPG synthesis. Despite its importance, the septal PGTase activity of FtsW 24 has not been documented in vivo. How its activity is spatiotemporally regulated in vivo has also 25 remained unknown. Here we investigated the septal PGTase activity and dynamics of FtsW in E. 26 coli cells using a combination of single-molecule imaging and genetic manipulations. We showed 27 that FtsW exhibited robust activity to incorporate an N-acetylmuramic acid analog at septa in the 28 absence of other known PGTases, confirming FtsW as the essential septum-specific PGTase in vivo. 29 Furthermore, we identified two populations of processively moving FtsW molecules at septa. A fast-30 moving population is driven by the treadmilling dynamics of FtsZ and independent of sPG 31 synthesis. A slow-moving population is driven by active sPG synthesis and independent of FtsZ 32treadmilling dynamics. We further identified that FtsN, a potential sPG synthesis activator, plays 33 an important role in promoting the slow-moving, sPG synthesis-dependent population. Our results 34 support a two-track model, in which inactive sPG synthase molecules follow the fast treadmilling 35 "Z-track" to be distributed along the septum; FtsN promotes their release from the "Z-track" to 36 become active in sPG synthesis on the slow "sPG-track". This model integrates spatial information 37 into the regulation of sPG synthesis activity and could serve as a mechanism for the spatiotemporal 38 coordination of bacterial cell wall constriction. 39To investigate the role of FtsW in sPG synthesis in vivo, we employed a cysteine-modification 40 inactivation assay 1,2 . Based on the homology structure of RodA 3 , we generated twelve ftsW C alleles that 41

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Section: Resultssupporting

confidence: 89%

Section: Resultsmentioning

confidence: 91%

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FtsW exhibits distinct processive motions driven by either septal cell wall synthesis or FtsZ treadmilling in E. coli

Yang¹,

McQuillen

Lyu

et al. 2019

Preprint

View full text Add to dashboard Cite

show abstract

“…Note that we do not model explicitly the stochastic on-reactions and off-reactions of individual subunits of an FtsZ filament, because such stochasticity is reflected in the treadmilling speed distribution of the filaments. The FtsZ filament length is set at 50 monomers (250 nm) and the treadmilling speed is independent of the filament length, in accordance to previous biochemical studies and a recent in vivo study 20,[32][33][34][35] . To discern principal interactions, the model considers one FtsI molecule and one FtsZ filament in a selfcontained septal section.…”

Section: Resultsmentioning

confidence: 84%

Treadmilling FtsZ polymers drive the directional movement of sPG-synthesis enzymes via a Brownian ratchet mechanism

McCausland¹,

Yang²,

Squyres

et al. 2021

Nat Commun

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The FtsZ protein is a central component of the bacterial cell division machinery. It polymerizes at mid-cell and recruits more than 30 proteins to assemble into a macromolecular complex to direct cell wall constriction. FtsZ polymers exhibit treadmilling dynamics, driving the processive movement of enzymes that synthesize septal peptidoglycan (sPG). Here, we combine theoretical modelling with single-molecule imaging of live bacterial cells to show that FtsZ’s treadmilling drives the directional movement of sPG enzymes via a Brownian ratchet mechanism. The processivity of the directional movement depends on the binding potential between FtsZ and the sPG enzyme, and on a balance between the enzyme’s diffusion and FtsZ’s treadmilling speed. We propose that this interplay may provide a mechanism to control the spatiotemporal distribution of active sPG enzymes, explaining the distinct roles of FtsZ treadmilling in modulating cell wall constriction rate observed in different bacteria.

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“…This process depends on FtsZ treadmilling and the activity of FtsZ-binding proteins, such as SepF, FtsA, ZapA or GpsB, which support filament formation, bundling, stabilization or membrane-anchoring (Eswara et al, 2018; Monteiro et al, 2018; Silber et al, 2020; Squyres et al, 2020; Whitley et al, 2020; Woldemeskel et al, 2017). Interestingly, our in vitro studies suggest that the SepH homologs from S. venezuelae and M. smegmatis display biochemical properties that are partially similar to the activities described for ZapA and GspB (Caldas et al, 2019; Eswara et al, 2018; Squyres et al, 2020; Woldemeskel et al, 2017). For example, similar to the effect described for GpsB, we found that SepH and SepH Ms stimulate FtsZ assembly (Eswara et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

A conserved cell division protein directly regulates FtsZ dynamics in filamentous and unicellular actinobacteria

Ramos-León

Bush

Sallmen

et al. 2020

Preprint

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Bacterial cell division is driven by the polymerization of the GTPase FtsZ into a contractile structure, the so-called Z-ring. This essential process involves proteins that modulate FtsZ dynamics and hence the overall Z-ring architecture. Actinobacteria, like Streptomyces and Mycobacterium lack known key FtsZ-regulators. Here we report the identification of SepH, a conserved actinobacterial protein that directly regulates FtsZ dynamics. We show that SepH is crucially involved in cell division in Streptomyces and that it binds FtsZ via a conserved helix-turn-helix motif, stimulating the assembly of FtsZ protofilaments. Comparative in vitro studies using the SepH homolog from Mycobacterium further reveal that SepH can also bundle FtsZ protofilaments, indicating an additional Z-ring stabilizing function in vivo. We propose that SepH plays a crucial role at the onset of cytokinesis in actinobacteria by promoting the rapid assembly of FtsZ filaments into division-competent Z-rings that can go on to mediate septum synthesis.

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Dynamics of bacterial cell division: Z ring condensation is essential for cytokinesis

Cited by 10 publications

References 51 publications

FtsW exhibits distinct processive motions driven by either septal cell wall synthesis or FtsZ treadmilling in E. coli

FtsW exhibits distinct processive motions driven by either septal cell wall synthesis or FtsZ treadmilling in E. coli

Treadmilling FtsZ polymers drive the directional movement of sPG-synthesis enzymes via a Brownian ratchet mechanism

A conserved cell division protein directly regulates FtsZ dynamics in filamentous and unicellular actinobacteria

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