In vitro assembly studies of FtsZ/tubulin-like proteins (TubZ) from Bacillus plasmids: evidence for a capping mechanism.
Proteins with a weak sequence similarity to tubulin and FtsZ are expressed from large plasmids of Bacillus anthracis and Bacillus thuringiensis and are probably involved in plasmid segregation. Previously designated RepX and TubZ, we designate them here as TubZ-Ba and TubZ-Bt. We have expressed and purified the proteins for in vitro studies. TubZ-Ba and TubZ-Bt share only 21% amino acid identity, but they have remarkably similar biochemical properties. They both assemble into two-stranded filaments and larger bundles above a critical concentration, and they hydrolyze GTP at a very high rate, approximately 20 GTP min(-1) TubZ(-1). Assembly is also supported by GTPgammaS. A tiny amount of GTPgammaS stabilizes polymers assembled in GTP and inhibits the GTPase by a mechanism involving cooperativity. The nucleotide in the polymers is almost 100% GDP, which is similar to microtubules but very different from the 20-30% GDP in FtsZ polymers. This suggests that the TubZ polymers have a capping mechanism that may be related to the GTP cap that produces dynamic instability of microtubules.
Chen
Yaodong
Y
Department of Cell Biology, Duke University Medical Center, Durham, NC 27710, USA.
Erickson
Harold P
HP
D001408:Bacillus anthracis / Q000235:genetics / Q000378:metabolism*; D001413:Bacillus thuringiensis / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D029968:Escherichia coli Proteins / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D008854:Microscopy, Electron; D010957:Plasmids / Q000235:genetics*; D014404:Tubulin / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure
D001426:Bacterial Proteins; D003598:Cytoskeletal Proteins; D029968:Escherichia coli Proteins; C071525:FtsZ protein, Bacteria; D014404:Tubulin; D006160:Guanosine Triphosphate; D020558:GTP Phosphohydrolases