Reconstitution of Protein Dynamics Involved in Bacterial Cell Division

Abstract

Even simple cells like bacteria have precisely regulated cellular anatomies, which allow them to grow, divide and to respond to internal or external cues with high fidelity. How spatial and temporal intracellular organization in prokaryotic cells is achieved and maintained on the basis of locally interacting proteins still remains largely a mystery. Bulk biochemical assays with purified components and in vivo experiments help us to approach key cellular processes from two opposite ends, in terms of minimal and maximal complexity. However, to understand how cellular phenomena emerge, that are more than the sum of their parts, we have to assemble cellular subsystems step by step from the bottom up. Here, we review recent in vitro reconstitution experiments with proteins of the bacterial cell division machinery and illustrate how they help to shed light on fundamental cellular mechanisms that constitute spatiotemporal order and regulate cell division.

The PAR proteins: from molecular circuits to dynamic self-stabilizing cell polarity

“…Mechanistic insights into the dynamics of polarization have come from biophysical and imaging approaches that allow one to quantify PAR protein dynamics in living cells, and from mathematical models that attempt to explain these dynamics in terms of known interactions. Looking forward, the system is now ripe for approaches from optogenetics, synthetic biology and biochemical reconstitution (Chau et al, 2012;Loose et al, 2017;Witte et al, 2017).…”

Functional Modules of Minimal Cell Division for Synthetic Biology

“…[1] In general, cell division has three main functions: i) it is the core of reproduction, [2] ii) it supports growth and renewal of tissues in multicellular organisms, [3] and iii) it mediates the formation of gametes for reproduction of multicellular organisms. Living cells have developed precisely regulated cellular machineries that enable growth, chromosome replication and segregation, division as well as response to internal and external stimuli.…”

“…Living cells have developed precisely regulated cellular machineries that enable growth, chromosome replication and segregation, division as well as response to internal and external stimuli. [1,2,5] The knowledge ii) Segregation of the bulk of the chromosome iii) Separation of the ter regions While E. coli is the most studied model organism, the details of how it (and other bacteria) manage chromosome segregation remain incomplete. [4] However, the underlying mechanistic and physicochemical details of how cell division and DNA segregation are achieved remain incomplete, although many important molecular building blocks have been identified to date.…”

“…[62] It could be demonstrated that the MinDE reaction and diffusion system can determine the localization of membrane attached molecules in a spatiotemporal manner without specific molecular interactions in vitro. [2,69] [67] In further experiments, an adapted cDICE method was used to transfer purified MinD and MinE into giant unilamellar vesicles (GUVs).…”

Lessons from a Minimal Genome: What Are the Essential Organizing Principles of a Cell Built from Scratch?

“…Positioning copies of the replicated genome to opposite sides of aS ynCell implies that division itself must be positioned at mid-cell.T here are an umber of positive and negative regulators that positiont he divisionm achinery in bacteria (reviewed in Monahan et al [50] ). However,o nly one has been successfully reconstitutedi nanumber of cell-free setups:t he E. coli Min/ FtsZ system (reviewed in Loose et al [51] and in Mizuuchi and Vecchiarelli [52] ). The Min system acts on at ubulin-like GTPase highly conservedi nt he microbial world called FtsZ.…”