bioRxiv ScienceSearch

bioRxiv · 10.1101/342766

Regulation of FtsZ levels in Escherichia coli in slow growth conditions

Abstract

A key regulator of cell division in most walled bacteria is the FtsZ protein that assembles into protofilaments attached to the membrane at midcell. These dynamic protofilament assemblies, known as the Z-ring, act as a scaffold for more than two dozen proteins involved in synthesis of septal cell envelopes. What triggers the formation of the Z-ring during the cell cycle is poorly understood. In Escherichia coli model organism, the common view is that FtsZ concentration is constant throughout its doubling time and therefore regulation of assembly should be controlled by some yet to be identified protein-protein interactions. Here we show using quantitative analysis of newly developed fluorescent reporter that FtsZ concentration varies in a cell-cycle dependent manner in slow growth conditions and that upregulation of FtsZ synthesis correlates with the formation of the Z-ring. About 4-fold upregulation of FtsZ synthesis in the first half of the cell cycle is followed by its rapid degradation by ClpXP protease in the last 10% of the cell cycle. The initiation of rapid degradation coincides with dissociation of FtsZ from the septum. Altogether, our data indicate that the Z-ring formation in slow growth conditions in E. coli is controlled by a regulatory sequence where upregulation of an essential cell cycle factor is followed by its degradation.\n\nSignificanceFtsZ is the key regulator for bacterial cell division. It initiates division by forming a dynamic ring-like structure, the Z-ring, at the mid-cell. Here we show that, contrarily to the current paradigm, FtsZ concentration in Escherichia coli model organism varies throughout cell cycle in slow growth conditions. Faster FtsZ synthesis in the first half of the cell cycle is followed by its rapid degradation by ClpXP protease in the end of the cell cycle. Upregulation of FtsZ synthesis correlates with the formation of the Z-ring. Our data demonstrates that in slow growth E. coli cell division progresses according to paradigm where upregulation of essential cell cycle factor is followed by its degradation.

Explore related subjects

Keep this discovery

BibTeXRIS

Männik, J., Walker, B. E.. 2018-06-08. Regulation of FtsZ levels in Escherichia coli in slow growth conditions. https://doi.org/10.1101/342766

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Functional characterization of Rho GTPase activating proteins SYDE1 and SYDE2

The human genome encodes more than 60 proteins containing Rho GTPase activating protein (RhoGAP) domains, many of which remain understudied with respect to their target specificity and biological roles. SYDE1 and SYDE2 are two such orphan RhoGAPs, for which there are few studies characterizing their biochemical and cellular functions and conflicting reports identifying their cognate GTPases. We previously identified SYDE1 and SYDE2 in a screen for substrates of the c-Jun N-terminal kinases. Here, we show that SYDE1 and SYDE2 are preferentially phosphorylated by JNK1 relative to other mitogen-activated protein kinases (MAPKs) at sites proximal to a kinase docking region. Purified SYDE1 and SYDE2 are shown to have significant catalytic GAP activity toward RhoA, Rac1, and Cdc42. However, neither up- nor down-regulation of SYDE1/2 expression leads to detectable changes in bulk GTP loading of any of these GTPases. Nevertheless, we demonstrate that SYDE1 and SYDE2, in a partially GAP-dependent manner, increase cell spreading and number of focal adhesions, and promote more directionally persistent migration in HEK293 cells. Together, these findings establish SYDE1 and SYDE2 as robust JNK substrates with catalytic activity toward a set of Rho GTPases and reveal basic functions of SYDE1 and SYDE2 in regulating cell morphology, adhesion, and migration.

cell biology

The filopodial scaffold polyphosphate dictates cell adhesion-versus-invasion decisions

Inorganic polyphosphate (polyP) is an ancient polymer conserved across all life, serving cell type and location specific functions in every major compartment. Yet its role at the plasma membrane, where it accumulates to peak levels in many primary cells, is largely unknown. Here we identify polyP as a stabilizing component of filopodia, actin based membrane protrusions that govern cell adhesion, contact inhibition, and chemotaxis. Elevating cellular polyP increases filopodial stability and enhances cell adhesion, whereas reducing polyP accelerates filopodial disassembly and promotes cell migration. Mechanistically, we find that polyP acts as a structural filopodial scaffold, recruiting and organizing IRSp53, a membrane curvature inducing protein. We show that metastatic fibroblasts and breast cancer organoids carry markedly reduced and intracellularly reorganized polyP levels relative to their non transformed counterparts. Restoring endogenous polyP via lipid nanoparticle delivery suppresses their invasive phenotypes and reverses prometastatic gene expression signatures, implicating polyP as a primordial tumor suppressor.

cell biology

Mitochondrial transfer mediates metabolic communication between beta cells and islet macrophages

Pancreatic islet macrophages support islet homeostasis and adapt their metabolic program in response to environmental cues, including beta cell released factors. Intercellular mitochondrial transfer is a biological process that modulates cellular responses. To test whether beta cells, which are strongly secretory, transfer mitochondria to islet macrophages, we generated mice with beta cell-specific expression of mitochondrial GFP (PhAMfloxIns1Cre). We demonstrate that beta cells transfer mitochondria to islet macrophages in vivo and in vitro. Diabetogenic stressors did not alter the frequency of mitochondrial transfer and macrophages containing beta cell-derived GFP exhibit increased protein synthesis rates. RNA-seq identified upregulation of activity-regulated cytoskeleton associated protein (Arc) in macrophages receiving beta cell-derived mitochondria, while disruption of actin cytoskeleton dynamics prevented mitochondrial transfer. Together, these findings identify mitochondrial transfer as a previously unrecognized mechanism of beta cell-macrophage communication that may contribute to islet homeostasis and immune regulation.

cell biology