Recruitment of the ParG Segregation Protein to Different Affinity DNA Sites

Zampini, Massimiliano; Derome, Andrew; Bailey, Simon E. S.; Barillà, Daniela; Hayes, Finbarr

doi:10.1128/jb.01630-08

Cited by 11 publications

(47 citation statements)

References 44 publications

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“…A 466 bp DNA fragment containing the promoter region of the tubRZ-Bs operon was amplified from plasmid pXK with unlabelled primer # 71 and 59-biotinylated primer # 72. Footprinting reactions were performed as described by Zampini et al (2009). Briefly, reactions containing biotin-labelled DNA (5 nM) and TubR-Bs (0.1-2.0 mM) were mixed in 50 ml EMSA binding buffer and incubated for 30 min at room temperature.…”

Section: Methodsmentioning

confidence: 99%

A new tubRZ operon involved in the maintenance of the Bacillus sphaericus mosquitocidal plasmid pBsph

Zhao

et al. 2014

Microbiology

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A new tubRZ operon involved in the maintenance of the Bacillus sphaericus mosquitocidal plasmid pBsph 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 TubZBs are involved in the replication and probably in the partitioning of pBsph plasmid, increasing our understanding of the genetic particularity of TubZ systems.

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Section: Methodsmentioning

confidence: 99%

A new tubRZ operon involved in the maintenance of the Bacillus sphaericus mosquitocidal plasmid pBsph

Zhao

et al. 2014

Microbiology

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“…The parH centromere that is recognized by ParG comprises a set of degenerate tetramer motifs separated by AT-rich spacers (33). ParG also is a transcriptional repressor of the parF-parG cassette which is achieved by binding to a second array of tetramer boxes upstream of the genes (38). Like other RHH proteins, ParG recognizes DNA via the antiparallel ␤-strands that comprise the ribbon element.…”

Section: Discussionmentioning

confidence: 99%

“…Extensive sequence variability in the ribbon element permits diverse RHH proteins to interact with discrete DNA sequences (37). In the case of ParG, the ribbon recognizes an array of degenerate tetramer boxes interspersed by AT-rich spacers both in the parH centromere, and at the operator site during transcriptional repression of the parF-parG genes (33,38).…”

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confidence: 99%

Breaking and Restoring the Hydrophobic Core of a Centromere-binding Protein

Saeed

Jowitt

Warwicker

et al. 2015

Journal of Biological Chemistry

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Background: Accurate segregation of antibiotic resistance plasmids requires dedicated centromere-binding proteins. Results: Assembly of the hydrophobic core of the ParG centromere-binding protein encoded by multiresistance plasmid TP228 is governed by a triad of key amino acids. Conclusion: ParG retains functionality with certain substitutions of the core triad. Significance: The study provides valuable insights into how multiresistance plasmids are maintained in bacteria.

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“…This segregation stability was shown to require a type Ib partition system encoding two proteins, the 22-kDa ParF protein and the 8.6-kDa ParG protein (20,22). Subsequently, it was demonstrated that ParG functions as the CBP that binds cooperatively to the parH centromere site to form a partition complex, which recruits ParF (23,24). ParG plays a second critical role in partition by stimulating the ATPase activity of ParF.…”

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confidence: 99%

Structural Mechanism of ATP-induced Polymerization of the Partition Factor ParF

Schumacher

Barge

et al. 2012

Journal of Biological Chemistry

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Background: ParF and ParG mediate TP228 plasmid segregation. Results: ATP binding to ParF activates segregation, and ADP binding to ParF antagonizes its segregation function. ParF-ADP is monomeric, and ParF-ATP is dimeric. ParF dimers assemble into polymers. Conclusion: ParF-ATP dimers serve as building blocks for polymer assembly. Significance: ParF-ATP polymers provide a mechanism for plasmid segregation.

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Recruitment of the ParG Segregation Protein to Different Affinity DNA Sites

Cited by 11 publications

References 44 publications

A new tubRZ operon involved in the maintenance of the Bacillus sphaericus mosquitocidal plasmid pBsph

A new tubRZ operon involved in the maintenance of the Bacillus sphaericus mosquitocidal plasmid pBsph

Breaking and Restoring the Hydrophobic Core of a Centromere-binding Protein

Structural Mechanism of ATP-induced Polymerization of the Partition Factor ParF

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