The MinDE system is a generic spatial cue for membrane protein distribution in vitro

Abstract

The E. coli MinCDE system has become a paradigmatic reaction–diffusion system in biology. The membrane-bound ATPase MinD and ATPase-activating protein MinE oscillate between the cell poles followed by MinC, thus positioning the main division protein FtsZ at midcell. Here we report that these energy-consuming MinDE oscillations may play a role beyond constraining MinC/FtsZ localization. Using an in vitro reconstitution assay, we show that MinDE self-organization can spatially regulate a variety of functionally completely unrelated membrane proteins into patterns and gradients. By concentration waves sweeping over the membrane, they induce a direct net transport of tightly membrane-attached molecules. That the MinDE system can spatiotemporally control a much larger set of proteins than previously known, may constitute a MinC-independent pathway to division site selection and chromosome segregation. Moreover, the here described phenomenon of active transport through a traveling diffusion barrier may point to a general mechanism of spatiotemporal regulation in cells.

Cell cycle regulated DNA methyltransferase: fluorescent tracking of a DNA strand-separation mechanism and identification of the responsible protein motif

“…Each measurement and background was averaged, and statistical outliers were eliminated. Statistical outliers were defined based on the maximum signal for each scan that was less than the first quartile (Q1) or greater than the third quartile (Q3) ( 18 ). Five scans were taken, and discarded reads were not replaced.…”

Synthetic cell division via membrane-transforming molecular assemblies

“…This gradient was capable of localizing FtsZ-YFP-MTS filaments to the middle of the compartment [61]. Very recently, our lab has shown that—even in the absence of MinC—MinD and MinE can support the anticorrelated movement and oscillation of model membrane proteins, including mCherry fused to various membrane targeting sequences, lipid-anchored streptavidin and FtsZ-YFP-MTS [88]. Moreover, if the proteins are permanently anchored to the membrane, MinDE oscillations can localize them to the middle of a microcompartment [88].…”

“…Very recently, our lab has shown that—even in the absence of MinC—MinD and MinE can support the anticorrelated movement and oscillation of model membrane proteins, including mCherry fused to various membrane targeting sequences, lipid-anchored streptavidin and FtsZ-YFP-MTS [88]. Moreover, if the proteins are permanently anchored to the membrane, MinDE oscillations can localize them to the middle of a microcompartment [88]. This implies that MinD and MinE are sufficient to generate a generic cue for the localization of membrane proteins, which may also be relevant for simplified divisome localization machineries.…”

Functional Modules of Minimal Cell Division for Synthetic Biology

“…[65] Membrane bound MinD-ATP recruits MinC with a subsequent inhibitory effect on FtsZ-ring assembly. [67] In further experiments, an adapted cDICE method was used to transfer purified MinD and MinE into giant unilamellar vesicles (GUVs). [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.…”