Dynamic Interaction of the Escherichia coli Cell Division ZipA and FtsZ Proteins Evidenced in Nanodiscs

Víctor M. Hernández-Rocamora,Allen P. Minton,Concepción García,Germán Rivas,Miguel Vicente,Belén Reija,Carlos Alfonso,Silvia Zorrilla,Paolo Natale

doi:10.1074/jbc.m112.388959

Abstract

The full-length ZipA protein from Escherichia coli, one of the essential components of the division proto-ring that provides membrane tethering to the septation FtsZ protein, has been incorporated in single copy into nanodiscs formed by a membrane scaffold protein encircling an E. coli phospholipid mixture. This is an acellular system that reproduces the assembly of part of the cell division components. ZipA contained in nanodiscs (Nd-ZipA) retains the ability to interact with FtsZ oligomers and with FtsZ polymers. Interactions with FtsZ occur at similar strengths as those involved in the binding of the soluble form of ZipA, lacking the transmembrane region, suggesting that the transmembrane region of ZipA has little influence on the formation of the ZipA·FtsZ complex. Peptides containing partial sequences of the C terminus of FtsZ compete with FtsZ polymers for binding to Nd-ZipA. The affinity of Nd-ZipA for the FtsZ polymer formed with GTP or GMPCPP (a slowly hydrolyzable analog of GTP) is moderate (micromolar range) and of similar magnitude as for FtsZ-GDP oligomers. Polymerization does not stabilize the binding of FtsZ to ZipA. This supports the role of ZipA as a passive anchoring device for the proto-ring with little implication, if any, in the regulation of its assembly. Furthermore, it indicates that the tethering of FtsZ to the membrane shows sufficient plasticity to allow for its release from noncentral regions of the cytoplasmic membrane and its subsequent relocation to midcell when demanded by the assembly of a division ring.

Highlights

ZipA provides membrane tethering to septation FtsZ protein
Using the nanodisc technology [22], we have investigated the interaction between the membrane-bound ZipA and the soluble FtsZ, two proteins that assemble in the E. coli proto-ring
In addition to their capacity to incorporate membrane proteins into a lipid environment while maintaining them in a soluble form, nanodiscs can be tagged with fluorescently labeled lipid molecules, which allowed us to use the full-length ZipA protein with no modification

Summary

Background

ZipA provides membrane tethering to septation FtsZ protein. Results: ZipA in nanodiscs moderately binds FtsZ oligomers and polymers . The full-length ZipA protein from Escherichia coli, one of the essential components of the division proto-ring that provides membrane tethering to the septation FtsZ protein, has been incorporated in single copy into nanodiscs formed by a membrane scaffold protein encircling an E. coli phospholipid mixture. This is an acellular system that reproduces the assembly of part of the cell division components. We describe here a synthetic system based on the integration of ZipA, one component of the division proto-ring, in nanodiscs These structures are formed by a membrane scaffold protein encircling a phospholipid bilayer [17]. We have measured the competition of two peptides derived from the C terminus of FtsZ in the binding of Nd-ZipA to FtsZ polymers

EXPERIMENTAL PROCEDURES

The abbreviations used are

RESULTS

DISCUSSION