Novel actin filaments from Bacillus thuringiensis form nanotubules for plasmid DNA segregation

Significance Actins and tubulins have dedicated functions that vary between eukaryotes and prokaryotes. During cell division, the prokaryotic contractile ring depends on the tubulin-like protein FtsZ, whereas this task relies on actin in eukaryotes. In contrast, microtubules orchestrate DNA segregation in eukaryotes, yet prokaryotic plasmid segregation often depends on actin-like proteins; this implies that actins and tubulins have somewhat interchangeable properties. Hence, we sought a bacterial filament that more closely resembles microtubules. Here, we report an actin from Bacillus thuringiensis that forms dynamic, antiparallel, two-stranded supercoiled filaments, which pair in the presence of a binding partner to form hollow cylinders. Thus, in this prokaryote, the actin fold has evolved to produce a filament system with comparable properties to the eukaryotic microtubule. Here we report the discovery of a bacterial DNA-segregating actin-like protein (BtParM) from Bacillus thuringiensis, which forms novel antiparallel, two-stranded, supercoiled, nonpolar helical filaments, as determined by electron microscopy. The BtParM filament features of supercoiling and forming antiparallel double-strands are unique within the actin fold superfamily, and entirely different to the straight, double-stranded, polar helical filaments of all other known ParMs and of eukaryotic F-actin. The BtParM polymers show dynamic assembly and subsequent disassembly in the presence of ATP. BtParR, the DNA-BtParM linking protein, stimulated ATP hydrolysis/phosphate release by BtParM and paired two supercoiled BtParM filaments to form a cylinder, comprised of four strands with inner and outer diameters of 57 Å and 145 Å, respectively. Thus, in this prokaryote, the actin fold has evolved to produce a filament system with comparable features to the eukaryotic chromosome-segregating microtubule.