Nano-encapsulated Escherichia coli Divisome Anchor ZipA, and in Complex with FtsZ

Sarah C. Lee, Richard Collins, Yu-pin Lin, Mohammed Jamshad, Claire Broughton, Sarah A. Harris, Benjamin S Hanson, Cecilia Tognoloni, Rosemary A. Parslow, Ann E. Terry, Alison Rodger, Corinne J. Smith, Karen J. Edler, Robert Ford, David I. Roper, Timothy R. Dafforn

Research output: Contribution to journalArticlepeer-review

16 Citations (SciVal)

Abstract

The E. coli membrane protein ZipA, binds to the tubulin homologue FtsZ, in the early stage of cell division. We isolated ZipA in a Styrene Maleic Acid lipid particle (SMALP) preserving its position and integrity with native E. coli membrane lipids. Direct binding of ZipA to FtsZ is demonstrated, including FtsZ fibre bundles decorated with ZipA. Using Cryo-Electron Microscopy, small-angle X-ray and neutron scattering, we determine the encapsulated-ZipA structure in isolation, and in complex with FtsZ to a resolution of 1.6 nm. Three regions can be identified from the structure which correspond to, SMALP encapsulated membrane and ZipA transmembrane helix, a separate short compact tether, and ZipA globular head which binds FtsZ. The complex extends 12 nm from the membrane in a compact structure, supported by mesoscale modelling techniques, measuring the movement and stiffness of the regions within ZipA provides molecular scale analysis and visualisation of the early divisome.
Original languageEnglish
Article number18712
JournalScientific Reports
Volume9
Issue number1
DOIs
Publication statusPublished - 10 Dec 2019

Funding

We would like to acknowledge the help of Ian Hands Portman in collection of negative stain transmission electron Microscopy images at the University of Warwick. This work is supported by BBSRC grant BB/J017310/1 (T.R.D, S.C.L, R.A.P). We would like to thank Diamond Light Source for the award of beamtime on instrument B21 (SM9727) and the assistance of Dr James Doutch in preparing the beamline and running this experiment. We would like to acknowledge ISIS Spallation Neutron and Muon Source for provision of neutron scattering facilities and the award of beamtime for experiment RB1410289 on SANS2D. We would also like to acknowledge Professor P. De Boer for providing the ZipA expression plasmid.

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