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Resolving new ultrastructural features of cytokinetic abscission with soft-X-ray cryo-tomography.


ABSTRACT: Mammalian cytokinetic abscission is mediated by the ESCRT membrane fission machinery. While much has been clarified on the topology and kinetics of abscission through high-resolution microscopy, key questions regarding the mechanism of abscission remain open. Here we apply cryogenic soft-X-ray tomography to elucidate new ultrastructural details in the intercellular membrane bridge connecting cells undergoing abscission. In particular, we resolve defined ring-like structures inside the midbody dark zone that have been inaccessible to EM, and identify membrane extrusions at the abscission sites. In cells at late stages of abscission we resolve a complex array of helical spirals, extending the structural information obtained by EM. Our results highlight the advantages of soft-X-ray tomography and emphasize the importance of using complementary approaches for characterizing cellular structures. Notably, by providing new structural data from intact cells we present a realistic view on the topology of abscission and suggest new mechanistic models for ESCRT mediated abscission.

SUBMITTER: Sherman S 

PROVIDER: S-EPMC4901327 | biostudies-other | 2016

REPOSITORIES: biostudies-other

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Resolving new ultrastructural features of cytokinetic abscission with soft-X-ray cryo-tomography.

Sherman Shachar S   Kirchenbuechler David D   Nachmias Dikla D   Tamir Adi A   Werner Stephan S   Elbaum Michael M   Elia Natalie N  

Scientific reports 20160610


Mammalian cytokinetic abscission is mediated by the ESCRT membrane fission machinery. While much has been clarified on the topology and kinetics of abscission through high-resolution microscopy, key questions regarding the mechanism of abscission remain open. Here we apply cryogenic soft-X-ray tomography to elucidate new ultrastructural details in the intercellular membrane bridge connecting cells undergoing abscission. In particular, we resolve defined ring-like structures inside the midbody da  ...[more]

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