title: Bacterial tubulin TubZ-Bt transitions between a two-stranded intermediate and a four-stranded filament upon GTP hydrolysis.
abstract: Cytoskeletal filaments form diverse superstructures that are highly adapted for specific functions. The recently discovered TubZ subfamily of tubulins is involved in type III plasmid partitioning systems, facilitating faithful segregation of low copy-number plasmids during bacterial cell division. One such protein, TubZ-Bt, is found on the large pBtoxis plasmid in Bacillus thuringiensis, and interacts via its extended C terminus with a DNA adaptor protein TubR. Here, we use cryo-electron microscopy to determine the structure of TubZ-Bt filaments and light scattering to explore their mechanism of polymerization. Surprisingly, we find that the helical filament architecture is remarkably sensitive to nucleotide state, changing from two-stranded to four-stranded depending on the ability of TubZ-Bt to hydrolyze GTP. We present pseudoatomic models of both the two- and four-protofilament forms based on cryo-electron microscopy reconstructions (10.8 Å and 6.9 Å, respectively) of filaments formed under different nucleotide states. These data lead to a model in which the two-stranded filament is a necessary intermediate along the pathway to formation of the four-stranded filament. Such nucleotide-directed structural polymorphism is to our knowledge an unprecedented mechanism for the formation of polar filaments.
authors.affiliation: Department of Biochemistry and Biophysics, and Howard Hughes Medical Institute, University of California, San Francisco, CA 94158.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Montabana
authors.forename: David A
authors.initials: DA
authors.lastname: Agard
mesh_terms: D001413:Bacillus thuringiensis / Q000737:chemistry*; D020285:Cryoelectron Microscopy; D018360:Crystallography, X-Ray; D004926:Escherichia coli; D006160:Guanosine Triphosphate / Q000378:metabolism*; D006868:Hydrolysis; D008958:Models, Molecular; D011487:Protein Conformation; D014404:Tubulin / Q000737:chemistry* / Q000378:metabolism
chemical_list: D014404:Tubulin; D006160:Guanosine Triphosphate
keywords: bacterial cytoskeleton; plasmid segregation