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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.

SUBMITTER: Holden SJ 

PROVIDER: S-EPMC3970495 | biostudies-literature | 2014 Mar

REPOSITORIES: biostudies-literature

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

Holden Seamus J SJ   Pengo Thomas T   Meibom Karin L KL   Fernandez Fernandez Carmen C   Collier Justine J   Manley Suliana S  

Proceedings of the National Academy of Sciences of the United States of America 20140310 12


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  ...[more]

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