PMID:21321206

Citation	Cho, H, McManus, HR, Dove, SL and Bernhardt, TG (2011) Nucleoid occlusion factor SlmA is a DNA-activated FtsZ polymerization antagonist.Proc. Natl. Acad. Sci. U.S.A. 108:3773-8
Abstract	The tubulin-like FtsZ protein initiates assembly of the bacterial cytokinetic machinery by polymerizing into a ring structure, the Z ring, at the prospective site of division. To block Z-ring formation over the nucleoid and help coordinate cell division with chromosome segregation, Escherichia coli employs the nucleoid-associated division inhibitor, SlmA. Here, we investigate the mechanism by which SlmA regulates FtsZ assembly. We show that SlmA disassembles FtsZ polymers in vitro. In addition, using chromatin immunoprecipitation (ChIP), we identified 24 SlmA-binding sequences (SBSs) on the chromosome. Remarkably, SlmA binding to SBSs dramatically enhanced its ability to interfere with FtsZ polymerization, and ChIP studies indicate that SlmA regulates FtsZ assembly at these sites in vivo. Because of the dynamic and highly organized nature of the chromosome, coupling SlmA activation to specific DNA binding provides a mechanism for the precise spatiotemporal control of its anti-FtsZ activity within the cell.
Links	PubMed Online version:10.1073/pnas.1018674108
Keywords
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Main Points of the Paper

• SlmA antagonizes FtsZ polymerization
• Two missense mutants are used
  • SlmA(R73D) is impaired in FtsZ binding
  • SlmA(T33A) is impaired in DNA binding
• DNA activates FtsZ's GTPase activity

Materials and Methods Used

• Plating assay
• Electrophoretic Mobility Shift Assay (EMSA)
• FtsZ Polymerization Assay
• Protein purification for FtsZ and SlmA
• SDS-PAGE
• Serial dilutions
• Fluorescence Microscopy
• Electron Microscopy
• Sedimentation Assay
• GTPase Assay

Phenotype Annotations

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Species	Taxon ID	Strain	Gene (if known)	OMP	Phenotype	Details	Evidence	Notes
Escherichia coli			slmA	absent DNA binding defective for nucleoid association	Other	missense mutation corresponding to T33A mutation in the protein	Binding Assay	Figure 2- EMSA
Escherichia coli			slmA	absent DNA binding defective for nucleoid association	Other	missense mutation corresponding to T33A mutation in the protein	Binding Assay	Figure 2- EMSA
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References

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