Colocalization of cell division proteins FtsZ and FtsA to cytoskeletal structures in living Escherichia coli cells by using green fluorescent protein.
In the current model for bacterial cell division, FtsZ protein forms a ring that marks the division plane, creating a cytoskeletal framework for the subsequent action of other proteins such as FtsA. This putative protein complex ultimately generates the division septum. Herein we report that FtsZ and FtsA proteins tagged with green fluorescent protein (GEP) colocalize to division-site ring-like structures in living bacterial cells in a visible space between the segregated nucleoids. Cells with higher levels of FtsZ-GFP or with FtsA-GFP plus excess wild-type FtsZ were inhibited for cell division and often exhibited bright fluorescent spiral tubules that spanned the length of the filamentous cells. This suggests that FtsZ may switch from a septation-competent localized ring to an unlocalized spiral under some conditions and that FtsA can bind to FtsZ in both conformations. FtsZ-GFP also formed nonproductive but localized aggregates at a higher concentration that could represent FtsZ nucleation sites. The general domain structure of FtsZ-GFP resembles that of tubulin, since the C terminus of FtsZ is not required for polymerization but may regulate polymerization state. The N-terminal portion of Rhizobium FtsZ polymerized in Escherichia coli and appeared to copolymerize with E. coli FtsZ, suggesting a degree of interspecies functional conservation. Analysis of several deletions of FtsA-GFP suggests that multiple segments of FtsA are important for its localization to the FtsZ ring.
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Department of Microbiology and Molecular Genetics, University of Texas Medical School, Houston 77030, USA.
Ehrhardt
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Margolin
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D000255:Adenosine Triphosphate / Q000378:metabolism; D001426:Bacterial Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000378:metabolism; D002455:Cell Division; D003598:Cytoskeletal Proteins; D003599:Cytoskeleton / Q000502:physiology* / Q000648:ultrastructure; D004926:Escherichia coli / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure; D029968:Escherichia coli Proteins; D019204:GTP-Binding Proteins / Q000032:analysis; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000032:analysis* / Q000096:biosynthesis; D010766:Phosphorylation; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis
D001426:Bacterial Proteins; D003598:Cytoskeletal Proteins; D029968:Escherichia coli Proteins; C065103:FtsA protein, Bacteria; C489310:FtsA protein, E coli; C071525:FtsZ protein, Bacteria; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D000255:Adenosine Triphosphate; D019204:GTP-Binding Proteins