title: High throughput 3D super-resolution microscopy reveals Caulobacter crescentus in vivo Z-ring organization.
abstract: We created a high-throughput modality of photoactivated localization microscopy (PALM) that enables automated 3D PALM imaging of hundreds of synchronized bacteria during all stages of the cell cycle. We used high-throughput PALM to investigate the nanoscale organization of the bacterial cell division protein FtsZ in live Caulobacter crescentus. We observed that FtsZ predominantly localizes as a patchy midcell band, and only rarely as a continuous ring, supporting a model of "Z-ring" organization whereby FtsZ protofilaments are randomly distributed within the band and interact only weakly. We found evidence for a previously unidentified period of rapid ring contraction in the final stages of the cell cycle. We also found that DNA damage resulted in production of high-density continuous Z-rings, which may obstruct cytokinesis. Our results provide a detailed quantitative picture of in vivo Z-ring organization.
authors.affiliation: Laboratory of Experimental Biophysics, École Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
authors.forename: Seamus J
authors.initials: SJ
authors.lastname: Holden
authors.forename: Thomas
authors.initials: T
authors.lastname: Pengo
authors.forename: Karin L
authors.initials: KL
authors.lastname: Meibom
authors.forename: Carmen
authors.initials: C
authors.lastname: Fernandez Fernandez
authors.forename: Justine
authors.initials: J
authors.lastname: Collier
authors.forename: Suliana
authors.initials: S
authors.lastname: Manley
mesh_terms: D016935:Caulobacter crescentus / Q000166:cytology* / Q000187:drug effects / Q000235:genetics; D002453:Cell Cycle; D004249:DNA Damage; D004269:DNA, Bacterial / Q000187:drug effects / Q000235:genetics; D008853:Microscopy / Q000379:methods*; D016685:Mitomycin / Q000494:pharmacology
chemical_list: D004269:DNA, Bacterial; D016685:Mitomycin
keywords: SOS response; bacterial cytoskeleton