3m8f
Protein structure of type III plasmid segregation TubR mutant
Structural highlights
FunctionTUBR_BACTI A DNA-binding protein that is part of the type III plasmid partition system used to ensure correct segregation of the pBtoxis plasmid. Cooperatively binds to the centromere-like site (tubC), which may seed filament formation by the TubZ polymerizing GTPase, stabilizing TubZ filaments. TubR-tubC complexes track the depolymerizing minus end of the filament, probably pulling plasmid within the cell (PubMed:20534443, PubMed:23010931, PubMed:25825718). Required for plasmid replication (PubMed:16936050, PubMed:17873046). Negatively regulates levels of TubZ; its effect on RNA expression has not been shown (Probable). Specifically binds iterons, 12-bp imperfect direct repeats that function as a plasmid origin of replication (PubMed:17873046, PubMed:23010931, PubMed:25825718). Four TubR dimers bind to tubC, forming an extended bent DNA-protein filament with protein wrapping helically around the outside of the DNA (PubMed:20534443, PubMed:23010931).[1] [2] [3] [4] [5] [6] Publication Abstract from PubMedThe segregation of plasmid DNA typically requires three elements: a DNA centromere site, an NTPase, and a centromere-binding protein. Because of their simplicity, plasmid partition systems represent tractable models to study the molecular basis of DNA segregation. Unlike eukaryotes, which utilize the GTPase tubulin to segregate DNA, the most common plasmid-encoded NTPases contain Walker-box and actin-like folds. Recently, a plasmid stability cassette on Bacillus thuringiensis pBtoxis encoding a putative FtsZ/tubulin-like NTPase called TubZ and DNA-binding protein called TubR has been described. How these proteins collaborate to impart plasmid stability, however, is unknown. Here we show that the TubR structure consists of an intertwined dimer with a winged helix-turn-helix (HTH) motif. Strikingly, however, the TubR recognition helices mediate dimerization, making canonical HTH-DNA interactions impossible. Mutagenesis data indicate that a basic patch, encompassing the two wing regions and the N termini of the recognition helices, mediates DNA binding, which indicates an unusual HTH-DNA interaction mode in which the N termini of the recognition helices insert into a single DNA groove and the wings into adjacent DNA grooves. The TubZ structure shows that it is as similar structurally to eukaryotic tubulin as it is to bacterial FtsZ. TubZ forms polymers with guanine nucleotide-binding characteristics and polymer dynamics similar to tubulin. Finally, we show that the exposed TubZ C-terminal region interacts with TubR-DNA, linking the TubR-bound pBtoxis to TubZ polymerization. The combined data suggest a mechanism for TubZ-polymer powered plasmid movement. Plasmid protein TubR uses a distinct mode of HTH-DNA binding and recruits the prokaryotic tubulin homolog TubZ to effect DNA partition.,Ni L, Xu W, Kumaraswami M, Schumacher MA Proc Natl Acad Sci U S A. 2010 Jun 4. PMID:20534443[7] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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