Cell-free biogenesis of bacterial division proto-rings that can constrict liposomes.
Journal
Communications biology
ISSN: 2399-3642
Titre abrégé: Commun Biol
Pays: England
ID NLM: 101719179
Informations de publication
Date de publication:
30 09 2020
30 09 2020
Historique:
received:
11
03
2020
accepted:
01
09
2020
entrez:
1
10
2020
pubmed:
2
10
2020
medline:
22
6
2021
Statut:
epublish
Résumé
A 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.
Identifiants
pubmed: 32999429
doi: 10.1038/s42003-020-01258-9
pii: 10.1038/s42003-020-01258-9
pmc: PMC7527988
doi:
Substances chimiques
Bacterial Proteins
0
Carrier Proteins
0
Cytoskeletal Proteins
0
Escherichia coli Proteins
0
FtsZ protein, Bacteria
0
Liposomes
0
ZapA protein, E coli
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
539Références
Shimizu, Y. et al. Cell-free translation reconstituted with purified components. Nat. Biotechnol. 19, 751–755 (2001).
pubmed: 11479568
doi: 10.1038/90802
Gros, J., Devbhandari, S. & Remus, D. Origin plasticity during budding yeast DNA replication in vitro. EMBO J. 33, 621–636 (2014).
pubmed: 24566988
pmcid: 3989655
doi: 10.1002/embj.201387278
Lee, K., Gallop, J. L., Rambani, K. & Kirschner, M. W. Self-Assembly of filopodia-like structures on supported lipid bilayers. Science 329, 1341–1345 (2010).
pubmed: 20829485
pmcid: 2982780
doi: 10.1126/science.1191710
Vignaud, T., Blanchoin, L. & Théry, M. Directed cytoskeleton self-organization. Trends Cell Biol. 22, 671–682 (2012).
pubmed: 23026031
doi: 10.1016/j.tcb.2012.08.012
Nguyen, P. A. et al. Spatial organization of cytokinesis signaling reconstituted in a cell-free system. Science 346, 244–247 (2014).
pubmed: 25301629
pmcid: 4281018
doi: 10.1126/science.1256773
Garner, E. C., Campbell, C. S., Weibel, D. B. & Mullins, R. D. Reconstitution of DNA segregation driven by assembly of a prokaryotic actin homolog. Science 315, 1270–1274 (2007).
pubmed: 17332412
pmcid: 2851738
doi: 10.1126/science.1138527
Dannhauser, P. N. & Ungewickell, E. J. Reconstitution of clathrin-coated bud and vesicle formation with minimal components. Nat. Cell Biol. 14, 634–639 (2012).
pubmed: 22522172
doi: 10.1038/ncb2478
Focus on the benefits of building life’s systems from scratch. Nature 563, 155–155 (2018).
Nourian, Z., Scott, A. & Danelon, C. Toward the assembly of a minimal divisome. Syst. Synth. Biol. 8, 237 (2014).
pubmed: 25136386
pmcid: 4127181
doi: 10.1007/s11693-014-9150-x
Scott, A. et al. Cell-free phospholipid biosynthesis by gene-encoded enzymes reconstituted in liposomes. PLoS ONE 11, e0163058 (2016).
pubmed: 27711229
pmcid: 5053487
doi: 10.1371/journal.pone.0163058
van Nies, P. et al. Self-replication of DNA by its encoded proteins in liposome-based synthetic cells. Nat. Commun. 9, 1583 (2018).
pubmed: 29679002
pmcid: 5910420
doi: 10.1038/s41467-018-03926-1
Godino, E. et al. De novo synthesized Min proteins drive oscillatory liposome deformation and regulate FtsA-FtsZ cytoskeletal patterns. Nat. Commun. 10, 4969 (2019).
pubmed: 31672986
pmcid: 6823393
doi: 10.1038/s41467-019-12932-w
Bi, E. & Lutkenhaus, J. FtsZ ring structure associated with division in Escherichia coli. Nature 354, 161–164 (1991).
pubmed: 1944597
doi: 10.1038/354161a0
Ma, X., Ehrhardt, D. W. & Margolin, W. Colocalization of cell division proteins FtsZ and FtsA to cytoskeletal structures in living Escherichia coli cells by using green fluorescent protein. Proc. Natl. Acad. Sci. 93, 12998–13003 (1996).
pubmed: 8917533
doi: 10.1073/pnas.93.23.12998
Hale, C. A. & de Boer, P. A. Recruitment of ZipA to the septal ring of Escherichia coli is dependent on FtsZ and independent of FtsA. J. Bacteriol. 181, 167–176 (1999).
pubmed: 9864327
pmcid: 103546
doi: 10.1128/JB.181.1.167-176.1999
Walker, B. E., Männik, J. & Mannik, J. Transient membrane-linked FtsZ assemblies precede Z-ring formation in Escherichia Coli. Curr. Biol. 30, 499–508 (2019).
doi: 10.1016/j.cub.2019.12.023
de Boer, P., Crossley, R. & Rothfield, L. The essential bacterial cell-division protein FtsZ is a GTPase. Nature 359, 254–256 (1992).
pubmed: 1528268
doi: 10.1038/359254a0
Mukherjee, A. & Lutkenhaus, J. Guanine nucleotide-dependent assembly of FtsZ into filaments. J. Bacteriol. 176, 2754–2758 (1994).
pubmed: 8169229
pmcid: 205420
doi: 10.1128/JB.176.9.2754-2758.1994
Pichoff, S. & Lutkenhaus, J. Unique and overlapping roles for ZipA and FtsA in septal ring assembly in Escherichia coli. EMBO J. 21, 685–693 (2002).
pubmed: 11847116
pmcid: 125861
doi: 10.1093/emboj/21.4.685
Pichoff, S. & Lutkenhaus, J. Tethering the Z ring to the membrane through a conserved membrane targeting sequence in FtsA. Mol. Microbiol. 55, 1722–1734 (2005).
pubmed: 15752196
doi: 10.1111/j.1365-2958.2005.04522.x
Hale, C. A. & de Boer, P. A. Direct binding of FtsZ to ZipA, an essential component of the septal ring structure that mediates cell division in E. coli. Cell 88, 175–185 (1997).
pubmed: 9008158
doi: 10.1016/S0092-8674(00)81838-3
Ortiz, C., Natale, P., Cueto, L. & Vicente, M. The keepers of the ring: regulators of FtsZ assembly. FEMS Microbiol. Rev. 40, 57–67 (2016).
pubmed: 26377318
doi: 10.1093/femsre/fuv040
Osawa, M. & Erickson, H. P. Liposome division by a simple bacterial division machinery. Proc. Natl Acad. Sci. USA 110, 11000–11004 (2013).
pubmed: 23776220
doi: 10.1073/pnas.1222254110
Cabré, E. J. et al. Bacterial division proteins FtsZ and ZipA induce vesicle shrinkage and cell membrane invagination. J. Biol. Chem. 288, 26625–26634 (2013).
pubmed: 23921390
pmcid: 3772209
doi: 10.1074/jbc.M113.491688
Ramirez-Diaz, D. A. et al. Treadmilling analysis reveals new insights into dynamic FtsZ ring architecture. PLOS Biol. 16, e2004845 (2018).
pubmed: 29775478
pmcid: 5979038
doi: 10.1371/journal.pbio.2004845
Krupka, M. et al. Escherichia coli FtsA forms lipid-bound minirings that antagonize lateral interactions between FtsZ protofilaments. Nat. Commun. 8, 15957 (2017).
pubmed: 28695917
pmcid: 5508204
doi: 10.1038/ncomms15957
Krupka, M., Sobrinos-Sanguino, M., Jiménez, M., Rivas, G. & Margolin, W. Escherichia coli ZipA organizes FtsZ Polymers into dynamic ring-like protofilament structures. MBio 9, e01008–18 (2018).
pubmed: 29921670
pmcid: 6016244
doi: 10.1128/mBio.01008-18
Loose, M. & Mitchison, T. J. The bacterial cell division proteins FtsA and FtsZ self-organize into dynamic cytoskeletal patterns. Nat. Cell Biol. 16, 38–46 (2014).
pubmed: 24316672
doi: 10.1038/ncb2885
Osawa, M., Anderson, D. E. & Erickson, H. P. Curved FtsZ protofilaments generate bending forces on liposome membranes. EMBO J. 28, 3476–3484 (2009).
pubmed: 19779463
pmcid: 2782090
doi: 10.1038/emboj.2009.277
Szwedziak, P., Wang, Q., Bharat, T. A. M., Tsim, M. & Löwe, J. Architecture of the ring formed by the tubulin homologue FtsZ in bacterial cell division. Elife 3, e04601 (2014).
pubmed: 25490152
pmcid: 4383033
doi: 10.7554/eLife.04601
Söderström, B. et al. Disassembly of the divisome in Escherichia coli: evidence that FtsZ dissociates before compartmentalization. Mol. Microbiol. 92, 1–9 (2014).
pubmed: 24506818
pmcid: 4004784
doi: 10.1111/mmi.12534
Daley, D. O., Skoglund, U. & Söderström, B. FtsZ does not initiate membrane constriction at the onset of division. Sci. Rep. 6, 33138 (2016).
pubmed: 27609565
pmcid: 5016829
doi: 10.1038/srep33138
Furusato, T. et al. De novo synthesis of basal bacterial cell division proteins FtsZ, FtsA, and ZipA inside giant vesicles. ACS Synth. Biol. 7, 953–961 (2018).
pubmed: 29510621
doi: 10.1021/acssynbio.7b00350
Doerr, A. et al. Modelling cell-free RNA and protein synthesis with minimal systems. Phys. Biol. 16, 025001 (2019).
pubmed: 30625117
doi: 10.1088/1478-3975/aaf33d
Blanken, D., van Nies, P. & Danelon, C. Quantitative imaging of gene-expressing liposomes reveals rare favorable phenotypes. Phys. Biol. 16, 045002 (2019).
pubmed: 30978176
doi: 10.1088/1478-3975/ab0c62
Rueda, S., Vicente, M. & Mingorance, J. Concentration and assembly of the division ring proteins FtsZ, FtsA, and ZipA during the Escherichia coli cell cycle. J. Bacteriol. 185, 3344–3351 (2003).
pubmed: 12754232
pmcid: 155373
doi: 10.1128/JB.185.11.3344-3351.2003
Caldas, P. et al. Cooperative ordering of treadmilling filaments in cytoskeletal networks of FtsZ and its crosslinker ZapA. Nat. Commun. 10, 5744 (2019).
pubmed: 31848350
pmcid: 6917738
doi: 10.1038/s41467-019-13702-4
Martos, A. et al. FtsZ polymers tethered to the membrane by ZipA are susceptible to spatial regulation by Min waves. Biophys. J. 108, 2371–2383 (2015).
pubmed: 25954894
pmcid: 4423045
doi: 10.1016/j.bpj.2015.03.031
Rivas, G., Alfonso, C., Jiménez, M., Monterroso, B. & Zorrilla, S. Macromolecular interactions of the bacterial division FtsZ protein: from quantitative biochemistry and crowding to reconstructing minimal divisomes in the test tube. Biophys. Rev. 5, 63–77 (2013).
pubmed: 28510160
pmcid: 5418439
doi: 10.1007/s12551-013-0115-1
Small, E. et al. FtsZ Polymer-bundling by the Escherichia coli ZapA orthologue, YgfE, involves a conformational change in cound GTP. J. Mol. Biol. 369, 210–221 (2007).
pubmed: 17428494
doi: 10.1016/j.jmb.2007.03.025
Martos, A. et al. Characterization of self-association and heteroassociation of bacterial cell division proteins FtsZ and ZipA in solution by composition gradient-static light scattering. Biochemistry 49, 10780–10787 (2010).
pubmed: 21082789
doi: 10.1021/bi101495x
Hernández-Rocamora, V. M. et al. Dynamic interaction of the Escherichia coli cell division ZipA and FtsZ proteins evidenced in nanodiscs. J. Biol. Chem. 287, 30097–30104 (2012).
pubmed: 22787144
pmcid: 3436366
doi: 10.1074/jbc.M112.388959
Osawa, M. & Erickson, H. P. Inside-out Z rings - constriction with and without GTP hydrolysis. Mol. Microbiol. 81, 571–579 (2011).
pubmed: 21631604
pmcid: 3229917
doi: 10.1111/j.1365-2958.2011.07716.x
Geissler, B., Elraheb, D. & Margolin, W. A gain-of-function mutation in ftsA bypasses the requirement for the essential cell division gene zipA in Escherichia coli. Proc. Natl. Acad. Sci. 100, 4197–4202 (2003).
pubmed: 12634424
doi: 10.1073/pnas.0635003100
Pichoff, S., Shen, B., Sullivan, B. & Lutkenhaus, J. FtsA mutants impaired for self-interaction bypass ZipA suggesting a model in which FtsA’s self-interaction competes with its ability to recruit downstream division proteins. Mol. Microbiol. 83, 151–167 (2012).
pubmed: 22111832
doi: 10.1111/j.1365-2958.2011.07923.x
Osawa, M., Anderson, D. E. & Erickson, H. P. Reconstitution of contractile FtsZ rings in liposomes. Science 320, 792–794 (2008).
pubmed: 18420899
pmcid: 2645864
doi: 10.1126/science.1154520
Cabré, E. J. et al. The nucleoid occlusion SlmA protein accelerates the disassembly of the FtsZ protein polymers without affecting their GTPase activity. PLoS ONE 10, e0126434 (2015).
pubmed: 25950808
pmcid: 4423959
doi: 10.1371/journal.pone.0126434
Monterroso, B. et al. The bacterial DNA binding protein MatP involved in linking the nucleoid terminal domain to the divisome at midcell interacts with lipid membranes. MBio 10, e00376–19 (2019).
pubmed: 31138739
pmcid: 6538776
doi: 10.1128/mBio.00376-19
Noireaux, V., Bar-Ziv, R. & Libchaber, A. Principles of cell-free genetic circuit assembly. Proc. Natl. Acad. Sci. 100, 12672–12677 (2003).
pubmed: 14559971
doi: 10.1073/pnas.2135496100
Mateos-Gil, P. et al. FtsZ polymers bound to lipid bilayers through ZipA form dynamic two dimensional networks. Biochim. Biophys. Acta 1818, 806–813 (2012).
pubmed: 22198391
doi: 10.1016/j.bbamem.2011.12.012
González, J. M. et al. Essential cell division protein FtsZ assembles into one monomer-thick ribbons under conditions resembling the crowded intracellular environment. J. Biol. Chem. 278, 37664–37671 (2003).
pubmed: 12807907
doi: 10.1074/jbc.M305230200
MacLean, B. et al. Skyline: an open source document editor for creating and analyzing targeted proteomics experiments. Bioinformatics 26, 966–968 (2010).
pubmed: 2844992
pmcid: 2844992
doi: 10.1093/bioinformatics/btq054
Schneider, C. A., Rasband, W. S. & Eliceiri, K. W. NIH Image to ImageJ: 25 years of image analysis. Nat. Methods 9, 671–675 (2012).
pubmed: 22930834
pmcid: 22930834
Rueden, C. T. et al. ImageJ2: ImageJ for the next generation of scientific image data. BMC Bioinformatics 18, 529 (2017).
pubmed: 29187165
pmcid: 5708080
doi: 10.1186/s12859-017-1934-z
Ball, G. et al. SIMcheck: a toolbox for successful super-resolution structured illumination microscopy. Sci. Rep. 5, 15915 (2015).
pubmed: 26525406
pmcid: 4648340
doi: 10.1038/srep15915