Helical Tubes of FtsZ from Methanococcus jannaschii

Abstract

Bacterial cell division depends on the formation of a cytokinetic ring structure, the Z-ring. The bacterial tubulin homologue FtsZ is required for Z-ring formation. FtsZ assembles into various polymeric forms in vitro, indicating a structural role in the septum of bacteria. We have used recombinant FtsZ1 protein from M. jannaschii to produce helical tubes and sheets with high yield using the GTP analogue GMPCPP [guanylyl-(α,β)-methylene-diphosphate]. The sheets appear identical to the previously reported Ca⁺⁺-induced sheets of FtsZ from M. jannaschii that were shown to consist of ‘thick’-filaments in which two protofilaments run in parallel. Tubes assembled either in Ca⁺⁺ or in GMPCPP contain filaments whose dimensions indicate that they could be equivalent to the ‘thick’-filaments in sheets. Some tubes are hollow but others are filled by additional protein density. Helical FtsZ tubes differ from eukaryotic microtubules in that the filaments curve around the filament axis with a pitch of ~ 430 Å for Ca⁺⁺-induced tubes or 590–620 Å for GMPCPP. However, their assembly in vitro as well-ordered polymers over distances comparable to the inner circumference of a bacterium may indicate a role in vivo. Their size and stability make them suitable for use in motility assays.