ZapE Is a Novel Cell Division Protein Interacting with FtsZ and Modulating the Z-Ring Dynamics

Marteyn, Benoît; Karimova, Gouzel; Fenton, Andrew K.; Gazi, Anastasia D.; West, Nicholas P.; Touqui, Lhousseine; Prévost, Marie‐Christine; Betton, Jean‐Michel; Poyraz, Ömer; Ladant, Daniel; Gerdes, Kenn; Sansonetti, Philippe J.; Tang, Christoph M.

doi:10.1128/mbio.00022-14

Cited by 52 publications

(74 citation statements)

References 28 publications

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“…The mla system, for maintaining outer membrane LPS integrity, and the poorly characterized membrane organizational protein, AsmA, were also important in vivo. Finally, a number of genes involved in cell division were intolerant of mutation in vivo; these may play a role in determining appropriate conditions for cellular replication, the regulation of which is likely crucial under stressful conditions (e.g., for zapE) (36).…”

Section: Significancementioning

confidence: 99%

Genome-wide screen identifies host colonization determinants in a bacterial gut symbiont

Powell

Leonard

Kwong

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

125

152

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Animal guts are often colonized by host-specialized bacterial species to the exclusion of other transient microorganisms, but the genetic basis of colonization ability is largely unknown. The bacterium Snodgrassella alvi is a dominant gut symbiont in honey bees, specialized in colonizing the hindgut epithelium. We developed methods for transposon-based mutagenesis in S. alvi and, using high-throughput DNA sequencing, screened genome-wide transposon insertion (Tnseq) and transcriptome (RNA-seq) libraries to characterize both the essential genome and the genes facilitating host colonization. Comparison of Tn-seq results from laboratory cultures and from monoinoculated worker bees reveal that 519 of 2,226 protein-coding genes in S. alvi are essential in culture, whereas 399 are not essential but are beneficial for gut colonization. Genes facilitating colonization fall into three broad functional categories: extracellular interactions, metabolism, and stress responses. Extracellular components with strong fitness benefits in vivo include trimeric autotransporter adhesins, O antigens, and type IV pili (T4P). Experiments with T4P mutants establish that T4P in S. alvi likely function in attachment and biofilm formation, with knockouts experiencing a competitive disadvantage in vivo. Metabolic processes promoting colonization include essential amino acid biosynthesis and iron acquisition pathways, implying nutrient scarcity within the hindgut environment. Mechanisms to deal with various stressors, such as for the repair of double-stranded DNA breaks and protein quality control, are also critical in vivo. This genome-wide study identifies numerous genetic networks underlying colonization by a gut commensal in its native host environment, including some known from more targeted studies in other hostmicrobe symbioses.type IV pilus | transposon mutagenesis | microbiota | symbiosis | Neisseriaceae

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Section: Significancementioning

confidence: 99%

Genome-wide screen identifies host colonization determinants in a bacterial gut symbiont

Powell

Leonard

Kwong

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

125

152

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“…Therefore, the regulation of Z-ring formation occurs at the level of FtsZ filament assembly. FtsZ interacting proteins modulate FtsZ polymerization and thus play important roles in this process (4,5,(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27). In E. coli at least 24 proteins have been identified that promote the assembly/disassembly processes of FtsZ at midcell (28,29).…”

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confidence: 99%

“…Among the many FtsZ regulatory proteins are the FtsZ ringassociated (Zap) proteins, ZapA-E (13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27). With the exception of ZapE, these proteins are recruited early during cytokinesis and have overlapping functions in stabilizing Z-ring formation at the midcell (13-17, 20, 21).…”

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confidence: 99%

Structural and Functional Analyses Reveal Insights into the Molecular Properties of the Escherichia coli Z Ring Stabilizing Protein, ZapC

Schumacher

Zhang

Huang

et al. 2016

Journal of Biological Chemistry

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In Escherichia coli cell division is driven by the tubulin-like GTPase, FtsZ, which forms the cytokinetic Z-ring. The Z-ring serves as a dynamic platform for the assembly of the multiprotein divisome, which catalyzes membrane cleavage to create equal daughter cells. Several proteins effect FtsZ assembly, thereby providing spatiotemporal control over cell division. One important class of FtsZ interacting/regulatory proteins is the Z-ring-associated proteins, Zaps, which typically modulate Z-ring formation by increasing lateral interactions between FtsZ protofilaments. Strikingly, these Zap proteins show no discernable sequence similarity, suggesting that they likely harbor distinct structures and mechanisms. The 19.8-kDa ZapC in particular shows no homology to any known protein. To gain insight into ZapC function, we determined its structure to 2.15 Å and performed genetic and biochemical studies. ZapC is a monomer composed of two domains, an N-terminal ␣/␤ region and a C-terminal twisted ␤ barrel-like domain. The structure contains two pockets, one on each domain. The N-domain pocket is lined with residues previously implicated to be important for ZapC function as an FtsZ bundler. The adjacent C-domain pocket contains a hydrophobic center surrounded by conserved basic residues. Mutagenesis analyses indicate that this pocket is critical for FtsZ binding. An extensive FtsZ binding surface is consistent with the fact that, unlike many FtsZ regulators, ZapC binds the large FtsZ globular core rather than C-terminal tail, and the presence of two adjacent pockets suggests possible mechanisms for ZapC-mediated FtsZ bundling.In Escherichia coli, the bacterial tubulin-like GTPase, FtsZ, drives cell division. Unlike the microtubule structures formed by tubulin, FtsZ forms protofilaments that combine to create a ring-like structure at midcell called the Z-ring, which mediates cell division (1-5). FtsZ is an ancient and highly conserved protein that is responsible for cell division in most bacteria as well as many archaea, all chloroplasts, and the mitochondria of primitive eukaryotes (4). FtsZ is made up of five main domains: an N-terminal region, which is largely disordered, a globular core that contains the nucleotide binding site and the T7 synergy loop needed for nucleotide hydrolysis, a long linker of variable sequence and length (ϳ40 -257 residues), a short ϳ15-residue C-terminal tail (CTT) 3 that contains a highly conserved set of residues critical for the docking of several FtsZ interacting proteins, and a more recently defined C-terminal variable (CTV) region (4, 6). Linear protofilaments are created by GTP binding to the globular domains of FtsZ between different protomers (1,7,8). The dynamic assembly and disassembly of protofilaments resulting from cycles of GTP binding and hydrolysis leads to Z-ring remodeling that is thought to contribute to the constrictive force required for cell division (4, 9, 10).In E. coli the intracellular levels of FtsZ remain essentially the same throughout the cell cycle and exceed...

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“…These Zap proteins stabilize FtsZ filaments prior to cell division by slowing depolymerization, thereby inhibiting GTPase activity (17,19,21,22). It has recently been shown that a fifth Zap protein, ZapE, also functions during division; however, it is thought to have a function opposing that of ZapA, ZapB, ZapC, and ZapD, as it binds to and causes the dissociation of FtsZ filaments (23). Although individually Zap proteins are not essential for divisome assembly, it is thought that the collective role they play in stabilizing FtsZ filaments prior to cell division is critical.…”

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confidence: 99%

Structure and Mutational Analyses of Escherichia coli ZapD Reveal Charged Residues Involved in FtsZ Filament Bundling

et al. 2016

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Bacterial cell division is an essential and highly coordinated process. It requires the polymerization of the tubulin homologue FtsZ to form a dynamic ring (Z-ring) at midcell. Z-ring formation relies on a group of FtsZ-associated proteins (Zap) for stability throughout the process of division. In Escherichia coli, there are currently five Zap proteins (ZapA through ZapE), of which four (ZapA, ZapB, ZapC, and ZapD) are small soluble proteins that act to bind and bundle FtsZ filaments. In particular, ZapD forms a functional dimer and interacts with the C-terminal tail of FtsZ, but little is known about its structure and mechanism of action. Here, we present the crystal structure of Escherichia coli ZapD and show it forms a symmetrical dimer with centrally located ␣-helices flanked by ␤-sheet domains. Based on the structure of ZapD and its chemical cross-linking to FtsZ, we targeted nine charged ZapD residues for modification by site-directed mutagenesis. Using in vitro FtsZ sedimentation assays, we show that residues R56, R221, and R225 are important for bundling FtsZ filaments, while transmission electron microscopy revealed that altering these residues results in different FtsZ bundle morphology compared to those of filaments bundled with wild-type ZapD. ZapD residue R116 also showed altered FtsZ bundle morphology but levels of FtsZ bundling similar to that of wild-type ZapD. Together, these results reveal that ZapD residues R116, R221, and R225 likely participate in forming a positively charged binding pocket that is critical for bundling FtsZ filaments. IMPORTANCEZ-ring assembly underpins the formation of the essential cell division complex known as the divisome and is required for recruitment of downstream cell division proteins. ZapD is one of several proteins in E. coli that associates with the Z-ring to promote FtsZ bundling and aids in the overall fitness of the division process. In the present study, we describe the dimeric structure of E. coli ZapD and identify residues that are critical for FtsZ bundling. Together, these results advance our understanding about the formation and dynamics of the Z-ring prior to bacterial cell division. Bacterial cell division is an essential and complex process that requires the coordinated assembly of a multiprotein molecular machine termed the divisome. The divisome is responsible for constriction of the inner and outer membranes, synthesis of septal peptidoglycan, and subsequent septum formation. In Escherichia coli, divisome proteins are recruited in a hierarchical manner and can be divided into three main groups based on their order of assembly: (i) the proto-ring, (ii) early divisome proteins, and (iii) late divisome proteins (1, 2). The successful assembly of the divisome depends on the initial formation of the Z-ring, which is comprised of the 40-kDa bacterial tubulin homologue FtsZ. FtsZ assembles into filaments in a GTP-dependent manner and a headto-tail fashion (3-6). The filaments are then tethered to the membrane, forming the Z-ring. They act as the...

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ZapE Is a Novel Cell Division Protein Interacting with FtsZ and Modulating the Z-Ring Dynamics

Cited by 52 publications

References 28 publications

Genome-wide screen identifies host colonization determinants in a bacterial gut symbiont

Genome-wide screen identifies host colonization determinants in a bacterial gut symbiont

Structural and Functional Analyses Reveal Insights into the Molecular Properties of the Escherichia coli Z Ring Stabilizing Protein, ZapC

Structure and Mutational Analyses of Escherichia coli ZapD Reveal Charged Residues Involved in FtsZ Filament Bundling

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