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13 p.-7 fig. ; Bacterial cell division is driven by an FtsZ ring in which the FtsZ protein localizes at mid-cell and recruits other proteins, forming a divisome. In Escherichia coli, the first molecular assembly of the divisome, the proto-ring, is formed by the association of FtsZ polymers to the cytoplasmic membrane through the membrane-tethering FtsA and ZipA proteins. The MinCDE system plays a major role in the site selection of the division ring because these proteins oscillate from pole to pole in such a way that the concentration of the FtsZ-ring inhibitor, MinC, is minimal at the cell center, thus favoring FtsZ assembly in this region. We show that MinCDE drives the formation of waves of FtsZ polymers associated to bilayers by ZipA, which propagate as antiphase patterns with respect to those of Min as revealed by confocal fluorescence microscopy. The emergence of these FtsZ waves results from the displacement of FtsZ polymers from the vicinity of the membrane by MinCD, which efficiently competes with ZipA for the C-terminal region of FtsZ, a central hub for multiple interactions that are essential for division. The coupling between FtsZ polymers and Min is enhanced at higher surface densities of ZipA or in the presence of crowding agents that favor the accumulation of FtsZ polymers near the membrane. The association of FtsZ polymers to the membrane modifies the response of FtsZ to Min, and comigrating Min-FtsZ waves are observed when FtsZ is free in solution and not attached to the membrane by ZipA. Taken together, our findings show that the dynamic Min patterns modulate the spatial distribution of FtsZ polymers in controlled minimal membranes. We propose that ZipA plays an important role in mid-cell recruitment of FtsZ orchestrated by MinCDE. ; This work was supported in part by the Human Frontiers Science Program through grant RGP0050/2010-C102 (to P.S. and G.R.), the DFG Leibniz Prize (to P.S.), the European Commission through contract HEALTH-F3-2009-223431 (to G.R.), and the Spanish Government through grant BIO2011-28941-C03-03 (to G.R.). ; Peer reviewed

20 p.-6 fig. ; FtsZ, the primary protein of the bacterial Z ring guiding cell division, has been recently shown to engage in intriguing treadmilling dynamics along the circumference of the division plane. When coreconstituted in vitro with FtsA, one of its natural membrane anchors, on flat supported membranes, these proteins assemble into dynamic chiral vortices compatible with treadmilling of curved polar filaments. Replacing FtsA by a membrane-targeting sequence (mts) to FtsZ, we have discovered conditions for the formation of dynamic rings, showing that the phenomenon is intrinsic to FtsZ. Ring formation is only observed for a narrow range of protein concentrations at the bilayer, which is highly modulated by free Mg2+ and depends upon guanosine triphosphate (GTP) hydrolysis. Interestingly, the direction of rotation can be reversed by switching the mts from the C-terminus to the N-terminus of the protein, implying that the filament attachment must have a perpendicular component to both curvature and polarity. Remarkably, this chirality switch concurs with previously shown inward or outward membrane deformations by the respective FtsZ mutants. Our results lead us to suggest an intrinsic helicity of FtsZ filaments with more than one direction of curvature, supporting earlier hypotheses and experimental evidence. ; BMBF/MPG (grant number 031A359A MaxSynBio). The MaxSynBio consortium is jointly funded by the Federal Ministry of Education and Research of Germany and the Max Planck Society. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. German-Israeli Foundation (GIF) (grant number 1160-137.14/2011). to MF and PS. AR is funded through the GIF for Scientific Research and Development. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Graduate School of Quantitative Biosciences of the Ludwig Maximilians University. DR-D and DG-S are supported by a DFG fellowship through QBM. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. JM is supported by the Nanosystems Initiative Munich (NIM) excellence cluster and the International Max Planck Research School for Molecular Life Sciences (IMPRS). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Human Frontiers Science Program (grant number RGP0050/2010). to GR and PS. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Spanish Government (grant number BFU2016-75471-C2-1-P). to GR. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Israel Academy of Science and Humanities (grant number 1701/13). to MF. ; Peer reviewed

11 p.-6 fig. ; FtsZ is a key component in bacterial cell division, being the primary protein of the presumably contractile Z ring. In vivo and in vitro, it shows two distinctive features that could so far, however, not be mechanistically linked: self-organization into directionally treadmilling vortices on solid supported membranes, and shape deformation of flexible liposomes. In cells, circumferential treadmilling of FtsZ was shown to recruit septum-building enzymes, but an active force production remains elusive. To gain mechanistic understanding of FtsZ dependent membrane deformations and constriction, we design an in vitro assay based on soft lipid tubes pulled from FtsZ decorated giant lipid vesicles (GUVs) by optical tweezers. FtsZ filaments actively transform these tubes into spring-like structures, where GTPase activity promotes spring compression. Operating the optical tweezers in lateral vibration mode and assigning spring constants to FtsZ coated tubes, the directional forces that FtsZ-YFP-mts rings exert upon GTP hydrolysis can be estimated to be in the pN range. They are sufficient to induce membrane budding with constricting necks on both, giant vesicles and E.coli cells devoid of their cell walls. We hypothesize that these forces result from torsional stress in a GTPase activity dependent manner. ; This work was funded through MaxSynBio (MPG together with Federal Ministry of Education and Research of Germany) Grant Number 031A359A to P.S., and the Transregio CRC 174 by the DFG (Deutsche Forschungsgemeinschaft) to P.S. and M.B. Work at GR Lab was supported by Spanish Government Grants BFU2016-75471-C2-1-P and 2019AEP088. ; Peer reviewed