High throughput 3D super-resolution microscopy reveals Caulobacter crescentus in vivo Z-ring organization

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 divisio...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2014-03, Vol.111 (12), p.4566-4571
Main Authors: Holden, Seamus J., Pengo, Thomas, Meibom, Karin L., Fernandez, Carmen Fernandez, Collier, Justine, Manley, Suliana
Format: Article
Language:English
Subjects:
Bacteria
Bacterial proteins
Biochemistry
Biological Sciences
Caulobacter crescentus
Caulobacter crescentus - cytology
Caulobacter crescentus - drug effects
Caulobacter crescentus - genetics
Cell Cycle
Cell division
Circles
Cytokinesis
DNA Damage
DNA, Bacterial - drug effects
DNA, Bacterial - genetics
Genetic SOS response
Imaging
Microscopy
Microscopy - methods
Mitomycin - pharmacology
Spatial resolution
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Summary: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.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1313368111