title: A new tubRZ operon involved in the maintenance of the Bacillus sphaericus mosquitocidal plasmid pBsph.
abstract: pBsph is a mosquitocidal plasmid first identified from Bacillus sphaericus, encoding binary toxins (Bin toxins) that are highly toxic to mosquito larvae. This plasmid plays an important role in the maintenance and evolution of the bin genes in B. sphaericus. However, little is known about its replication and partitioning. Here, we identified a 2.4 kb minimal replicon of pBsph plasmid that contained an operon encoding TubR-Bs and TubZ-Bs, each of which was shown to be required for plasmid replication. TubR-Bs was shown to be a transcriptional repressor of tubRZ-Bs genes and could bind cooperatively to the replication origin of eleven 12 bp degenerate repeats in three blocks, and this binding was essential for plasmid replication. TubZ-Bs exhibited GTPase activities and interacted with TubR-Bs : DNA complex to form a ternary nucleoprotein apparatus. Electron and fluorescence microscopy revealed that TubZ-Bs assembled filaments both in vitro and in vivo, and two point mutations in TubZ-Bs (T114A and Y260A) that severely impaired the GTPase and polymerization activities were found to be defective for plasmid maintenance. Further investigation demonstrated that overproduction of TubZ-Bs-GFP or its mutant forms significantly reduced the stability of pBsph. Taken together, these results suggested that TubR-Bs and TubZ-Bs are involved in the replication and probably in the partitioning of pBsph plasmid, increasing our understanding of the genetic particularity of TubZ systems.
authors.affiliation: University of the Chinese Academy of Sciences, Beijing 100039, PR China.|Key Laboratory of Applied and Environmental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, PR China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Ge
authors.affiliation: Key Laboratory of Applied and Environmental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, PR China.
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Hu
authors.affiliation: Key Laboratory of Applied and Environmental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, PR China.
authors.forename: Ni
authors.initials: N
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Applied and Environmental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, PR China.
authors.forename: Tingyu
authors.initials: T
authors.lastname: Shi
authors.affiliation: Key Laboratory of Applied and Environmental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, PR China.
authors.forename: Quanxin
authors.initials: Q
authors.lastname: Cai
authors.affiliation: Key Laboratory of Applied and Environmental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, PR China.
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Yuan
mesh_terms: D001407:Bacillus / Q000235:genetics*; D001427:Bacterial Toxins / Q000235:genetics; D004261:DNA Replication; D005798:Genes, Bacterial; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D009876:Operon; D010957:Plasmids; D011485:Protein Binding; D014158:Transcription, Genetic
chemical_list: D001427:Bacterial Toxins