Measuring abscission spatiotemporal dynamics using quantitative high-resolution microscopy

Gershony, Ofir; Sherman, Shachar; Adar, Shai; Segal, Inbar; Nachmias, Dikla; Goliand, Inna; Elia, Natalie

doi:10.1016/bs.mcb.2016.03.032

Cited by 17 publications

(18 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To avoid over expression artifacts we immunolabeled the ESCRT-III component IST1 in Hela cells that were synchronized to enrich the percentage of cells undergoing abscission. Microtubule staining was performed for determining abscission progression as previously described (Elia et al, 2011;Gershony et al, 2017). In bridges at early stages, IST1 was found in two large diameter cortical rings residing at the rims of the dark zone ( Figure 1A, ring diameter 1.09±0.26 µm); consistent with previous data obtained for other ESCRT III proteins (Elia et al, 2011;Goliand et al, 2014).…”

Section: Resultssupporting

confidence: 85%

Resolving ESCRT-III spirals at the intercellular bridge of dividing cells using 3D STORM imaging

Goliand

Dadosh

Elia

2017

Preprint

View full text Add to dashboard Cite

Section: Resultssupporting

confidence: 85%

Resolving ESCRT-III spirals at the intercellular bridge of dividing cells using 3D STORM imaging

Goliand

Dadosh

Elia

2017

Preprint

View full text Add to dashboard Cite

“…We found that the second abscission usually occurred within 15 minutes of the first abscission (~75% of the cells), in both siLUC and siKIF-treated cells (Figure 7D). This is similar to but slightly faster than what was reported using GFP-tubulin (Guizetti et al, 2011;Gershony et al, 2017). Interestingly, the cells that had the longest times to first abscission were not the ones that had the longest times between first and second abscissions, suggesting they are independent events (data not shown).…”

Section: Kif20b Depletion Disrupts Abscission Timingsupporting

confidence: 85%

Kinesin-6 KIF20B is required for efficient cytokinetic furrowing and timely abscission in human cells

Janisch

McNeely

Dardick

et al. 2017

Preprint

View full text Add to dashboard Cite

Cytokinesis requires the cooperation of many cytoskeletal and membrane regulators. Most of the major players required for cytokinesis are known, but the temporal regulation and adaptations for different cell types are less understood. KIF20B (previously called MPHOSPH1 or MPP1) is a member of the Kinesin-6 family, which also includes the better-known members KIF23/MKLP1 and KIF20A/MKLP2. Previously, we showed that mouse Kif20b is involved in cerebral cortex growth and midbody organization of neural stem cells. Here we show that KIF20B has a cellautonomous role in cytokinesis in isolated human cells. It localizes to microtubules of the central spindle and midbody throughout cytokinesis, at sites distinct from the other Kinesin-6 family members. KIF20B is not required for central spindle or midbody assembly, but affects midbody shape and late maturation steps. KIF20B appears to temporally regulate both furrow ingression and abscission.

show abstract

“…We found antibodies for IST1, an ESCRT-III protein shown to be crucial for abscission and to polymerize into tubes in vitro (Bajorek et al, 2009;McCullough et al, 2015), most suitable to our study. Microtubules were stained for visualizing the intercellular bridge (Elia et al, 2011;Gershony et al, 2017).…”

Section: Resultsmentioning

confidence: 99%

“…A measure of abscission progression is the extent of bridge narrowing, which can be determined based on microtubule staining (Elia et al, 2011;Gershony et al, 2017). To substantiate the scenario described above, we measured the relative bridge constriction in each stage ( Figure 2C).…”

Section: Resultsmentioning

confidence: 99%

Resolving ESCRT-III Spirals at the Intercellular Bridge of Dividing Cells Using 3D STORM

et al. 2018

View full text Add to dashboard Cite

The ESCRT machinery mediates membrane fission in a variety of processes in cells. According to current models, ESCRT-III proteins drive membrane fission by assembling into helical filaments on membranes. Here, we used 3D STORM imaging of endogenous ESCRT-III component IST1 to reveal the evolution of the structural organization of ESCRT-III in mammalian cytokinetic abscission. Using this approach, ESCRT-III ring and spiral assemblies were resolved and characterized at different stages of abscission. Visualization of IST1 structures in cells lacking the microtubule-severing enzyme spastin and in cells depleted of specific ESCRT-III components or the ATPase VPS4 demonstrated the contribution of these components to the organization and function of ESCRTs in cells. This work provides direct evidence that ESCRT-III proteins form helical filaments to mediate their function in cells and raises new mechanistic scenarios for ESCRT-driven cytokinetic abscission.

show abstract

Measuring abscission spatiotemporal dynamics using quantitative high-resolution microscopy

Cited by 17 publications

References 21 publications

Resolving ESCRT-III spirals at the intercellular bridge of dividing cells using 3D STORM imaging

Resolving ESCRT-III spirals at the intercellular bridge of dividing cells using 3D STORM imaging

Kinesin-6 KIF20B is required for efficient cytokinetic furrowing and timely abscission in human cells

Resolving ESCRT-III Spirals at the Intercellular Bridge of Dividing Cells Using 3D STORM

Contact Info

Product

Resources

About