Caveolae promote successful abscission by controlling intercellular bridge tension during cytokinesis

Andrade, Virginia; Bai, Jing; Gupta-Rossi, Neetu; Jiménez, Ana Joaquina; Delevoye, Cédric; Lamaze, Christophe; Échard, Arnaud

doi:10.1126/sciadv.abm5095

Cited by 27 publications

(39 citation statements)

References 81 publications

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“…In midbody-containing cells, all three MIT proteins co-localized with CEP55 ring structures on either side of the Flemming body ( Figure 5A–C ). Uniquely, KATNA1 also distributed along the midbody arms, reminiscent of localization observed for caveolae, and termed ‘midbody entry points’ ( Andrade et al, 2022 ). Arm staining was particularly prominent in early midbody-stage cells ( Figure 5B ), and a similar KATNA1 staining pattern has been reported in rat cell lines ( Matsuo et al, 2013 ).…”

Section: Resultsmentioning

confidence: 96%

Comprehensive analysis of the human ESCRT-III-MIT domain interactome reveals new cofactors for cytokinetic abscission

Wenzel

Mackay

Skalicky

et al. 2022

eLife

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The 12 related human ESCRT-III proteins form filaments that constrict membranes and mediate fission, including during cytokinetic abscission. The C-terminal tails of polymerized ESCRT-III subunits also bind proteins that contain Microtubule-Interacting and Trafficking (MIT) domains. MIT domains can interact with ESCRT-III tails in many different ways to create a complex binding code that is used to recruit essential cofactors to sites of ESCRT activity. Here, we have comprehensively and quantitatively mapped the interactions between all known ESCRT-III tails and 19 recombinant human MIT domains. We measured 228 pairwise interactions, quantified 60 positive interactions, and discovered 18 previously unreported interactions. We also report the crystal structure of the SPASTIN MIT domain in complex with the IST1 C-terminal tail. Three MIT enzymes were studied in detail and shown to: (1) localize to cytokinetic midbody membrane bridges through interactions with their specific ESCRT-III binding partners (SPASTIN-IST1, KATNA1-CHMP3, and CAPN7-IST1), (2) function in abscission (SPASTIN, KATNA1, and CAPN7), and (3) function in the ‘NoCut’ abscission checkpoint (SPASTIN and CAPN7). Our studies define the human MIT-ESCRT-III interactome, identify new factors and activities required for cytokinetic abscission and its regulation, and provide a platform for analyzing ESCRT-III and MIT cofactor interactions in all ESCRT-mediated processes.

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Section: Resultsmentioning

confidence: 96%

Comprehensive analysis of the human ESCRT-III-MIT domain interactome reveals new cofactors for cytokinetic abscission

Wenzel

Mackay

Skalicky

et al. 2022

eLife

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“…First, in contrast to the stereotypical CB that connects the soma of two cells and aligns predominantly with the nuclei of daughter cells (Andrade et al, 2022), some of our ICs connected to other parts of the cell, at times even connecting a process of one cell to the soma of the other, rather than simply connecting soma to soma in an aligned fashion (Fig. 3A).…”

Section: Morphological Features Of Connected Gcs Demonstrate Structur...mentioning

confidence: 78%

“…One of the most peculiar features observed within ICs were electron-dense regions, which we presumed to be microtubule bundles (Andrade et al, 2022;Zenker et al, 2017). We quantified the overall voxel intensity in each cross-section along ICs as a proxy for cytoskeletal filament density (Fig.…”

Section: Ics Are Anatomically Diverse and Complexmentioning

confidence: 99%

“…1C-E'; Movie S2). CBs are microtubule-rich connections that play a vital role in the abscission of dividing cells; however, observations of CBs have been limited to in vitro models (Andrade et al, 2022;Guizetti et al, 2011). Full segmentation of an IC revealed two features previously ascribed to CBs: (1) an electron-dense structure within the tube, which could indicate the presence of a midbody (MB); and (2) microtubules emanating from the dense region into both cells (Fig.…”

Section: Identification Of Cerebellar Granule Cells Linked By Interce...mentioning

confidence: 99%

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3D reconstruction of the cerebellar germinal layer reveals intercytoplasmic connections between developing granule cells

Cervantes

Khare

Wilson

et al. 2022

Preprint

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The difficulty of retrieving high-resolution, in vivo evidence of the proliferative- and migratory processes occurring in neural germinal zones has limited our understanding of neurodevelopmental mechanisms. Here, we employed a connectomic approach using a high-resolution, serial-sectioning scanning electron microscopy volume to investigate the laminar cytoarchitecture of the transient external granular layer (EGL) of the developing cerebellum, where granule cells coordinate a series of mitotic and migratory events. By integrating image segmentation, 3D reconstruction, and deep learning approaches, we discovered and characterized anatomically complex intercellular connections bridging pairs of cerebellar granule cells throughout the EGL. Connected cells were either mitotic, migratory, or transitioning between these two cell stages, displaying a chronological continuum of proliferative and migratory events never previously observed in vivo at this resolution. This unprecedented ultra-structural characterization poses intriguing hypotheses about intercellular connectivity between developing progenitors, and its possible role in the development of the central nervous system (CNS).

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“…4 A), a region where the plasma membrane is rather curved and reported in E. coli to be enriched in cardiolipin [ 42 , 45 , 46 ]. The fact that these vesicles were observed at the septum of dividing bacteria is reminiscent of a similar localization for eukaryotic caveolae in non-transfected organisms, where caveolin and caveolae are involved in membrane changes during abscission and cytokinesis [ 47 ]. Finally, this situation of MP-lipid relationships is possibly an illustration of the important role of cardiolipin that has been involved for intracellular curved membrane structures formation in E. coli when overexpressing some specific MPs [ 48 , 49 ].…”

Section: Discussionmentioning

confidence: 99%

Formation of intracellular vesicles within the Gram+ Lactococcus lactis induced by the overexpression of Caveolin-1β

et al. 2022

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Background Caveolae are invaginated plasma membrane domains of 50–100 nm in diameter involved in many important physiological functions in eukaryotic cells. They are composed of different proteins, including the membrane-embedded caveolins and the peripheric cavins. Caveolin-1 has already been expressed in various expression systems (E. coli, insect cells, Toxoplasma gondii, cell-free system), generating intracellular caveolin-enriched vesicles in E. coli, insect cells and T. gondii. These systems helped to understand the protein insertion within the membrane and its oligomerization. There is still need for fundamental insights into the formation of specific domains on membrane, the deformation of a biological membrane driven by caveolin-1, the organization of a caveolar coat, and the requirement of specific lipids and proteins during the process. The aim of this study was to test whether the heterologously expressed caveolin-1β was able to induce the formation of intracellular vesicles within a Gram+ bacterium, Lactococcus lactis, since it displays a specific lipid composition different from E. coli and appears to emerge as a good alternative to E. coli for efficient overexpression of various membrane proteins. Results Recombinant bacteria transformed with the plasmid pNZ-HTC coding for the canine isoform of caveolin-1β were shown to produce caveolin-1β, in its functional oligomeric form, at a high expression level unexpected for an eukaryotic membrane protein. Electron microscopy revealed several intracellular vesicles from 30 to 60 nm, a size comparable to E. coli h-caveolae, beneath the plasma membrane of the overexpressing bacteria, showing that caveolin-1β is sufficient to induce membrane vesiculation. Immunolabelling studies showed antibodies on such neo-formed intracellular vesicles, but none on plasma membrane. Density gradient fractionation allowed the correlation between detection of oligomers on Western blot and appearance of vesicles measurable by DLS, showing the requirement of caveolin-1β oligomerization for vesicle formation. Conclusions Lactococcus lactis cells can heterologously overexpress caveolin-1β, generating caveolin-1β enriched intracellular neo-formed vesicles. These vesicles might be useful for potential co-expression of membrane proteins of pharmaceutical interest for their simplified functional characterization.

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Caveolae promote successful abscission by controlling intercellular bridge tension during cytokinesis

Cited by 27 publications

References 81 publications

Comprehensive analysis of the human ESCRT-III-MIT domain interactome reveals new cofactors for cytokinetic abscission

Comprehensive analysis of the human ESCRT-III-MIT domain interactome reveals new cofactors for cytokinetic abscission

3D reconstruction of the cerebellar germinal layer reveals intercytoplasmic connections between developing granule cells

Formation of intracellular vesicles within the Gram+ Lactococcus lactis induced by the overexpression of Caveolin-1β

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