Resolving new ultrastructural features of cytokinetic abscission with soft-X-ray cryo-tomography.
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.
Department of Life Sciences and NIBN, Ben Gurion University of the Negev, Beer Sheva, Israel.
Shachar
S
Sherman
Department of Materials and Interfaces, Weizmann Institute of Science, Rehovot, Israel.
David
D
Kirchenbuechler
Department of Life Sciences and NIBN, Ben Gurion University of the Negev, Beer Sheva, Israel.
Dikla
D
Nachmias
Department of Life Sciences and NIBN, Ben Gurion University of the Negev, Beer Sheva, Israel.
Adi
A
Tamir
Helmholtz Zentrum Berlin für Materialien und Energie GmbH, Wilhelm-Conrad-Röntgen Campus, Berlin, Germany.
Stephan
S
Werner
Department of Materials and Interfaces, Weizmann Institute of Science, Rehovot, Israel.
Michael
M
Elbaum
Department of Life Sciences and NIBN, Ben Gurion University of the Negev, Beer Sheva, Israel.
Natalie
N
Elia