Biology subjects

Plastino, J.

Publications and source records attributed to Plastino, J..

4 recordsLinked to original sources

bioRxiv2025

The actin protrusion deforms the nucleus during invasion through basement membrane

Cell invasion through basement membrane (BM) extracellular matrix barriers is important during organ development, immune cell trafficking, and cancer metastasis. Here we study an invasion event, anchor cell (AC) invasion, which occurs during Caenorhabditis elegans development. The actin protrusion of the invading AC deforms then disrupts the BM. With live-cell imaging, we observe that the distal end of the actin protrusion contacts the nucleus during wild-type invasion, and deforms the nuclear envelope. Further we show that there is a correlation between invasion efficiency and nuclear contact: under mutant conditions where invasion is reduced, nuclear deformation is diminished. Knocking down the nuclear cytoskeleton protein lamin produces a nuclear positioning defect in the AC and reduces invasion efficiency. In keeping with this, both actin and microtubule-associated nesprins, ANC-1 and UNC-83 respectively, are found on the AC nuclear envelope during invasion, and perturbing their function reduces invasion. CEC-4, a chromodomain protein that attaches chromatin to the nuclear envelope in C. elegans, is also found to promote AC invasion. All together these data indicate that the positioning and properties of the AC nucleus are important for BM invasion. SUMMARY STATEMENTActin-based membrane protrusions in invading cells apply force to basement membrane (BM) barriers to help break through them. Here we provide evidence that the nucleus also plays a role in optimizing BM invasion.

cell biology↗

bioRxiv2019

Actin accumulates nesprin-2 at the front of the nucleus during confined cell migration

The mechanisms by which cells exert forces on their nuclei to migrate through openings smaller than the nuclear diameter remain unclear. In microfluidic devices, the hourglass shape of the nucleus and its strain patterns as it translocates through narrow constrictions suggest pulling forces. We use CRISPR/Cas9 to fluorescently label nesprin-2 giant, a protein that links the cytoskeleton to the interior of the nucleus. We demonstrate that nesprin-2 giant accumulates at the front of the nucleus during nuclear deformation through narrow constrictions, independently of the nuclear lamina. We find that nesprins are more mobile than lamin A/C, at time scales similar to that of the accumulation. Using artificial constructs, we show that the actin-binding domain of nesprin-2 is necessary and sufficient to generate this accumulation, and that microtubules are not necessary. Actin filaments are organized in a barrel structure around the moving nucleus in the direction of movement, suggesting that this structure is responsible for redistribution of nesprins towards the front of the nucleus. Two-photon ablation and the use of drugs inhibiting the cytoskeleton demonstrate a pulling force on the nucleus from the front of the cell that is dependent on formin and actomyosin contractility. This elastic recoil is significantly reduced when nesprins are reduced at the nuclear envelope. We thus show that actin redistributes nesprin-2 giant towards the front of the nucleus and contributes to pulling the nucleus through narrow constrictions, in concert with myosin.

cell biology↗

bioRxiv2019

Actin modulates shape and mechanics of tubular membranes

The actin cytoskeleton shapes cells and also organizes internal membranous compartments. In particular, it interacts with membranes in intracellular transport of material in mammalian cells, yeast or plant cells. Tubular membrane intermediates, pulled along microtubule tracks, are involved during these processes, and destabilize into vesicles. While the role of actin in this destabilization process is still debated, literature also provide examples of membranous structures stabilization by actin. To directly address this apparent contradiction, we mimic the geometry of tubular intermediates with preformed membrane tubes. The growth of an actin sleeve at the tube surface is monitored spatio-temporally. Depending on network cohesiveness, actin is able to stabilize, or maintain membrane tubes under pulling. Indeed, on a single tube, thicker portions correlate with the presence of actin. Such structures relax over several minutes, and may provide enough time and curvature geometries for other proteins to act on tube stability.

biophysics↗

bioRxiv2018

A direct proof that sole actin dynamics drive membrane deformations

Cell membrane deformations are crucial for proper cell function. Specialized protein assemblies initiate inward or outward membrane deformations that turn into, for example, filopodia or endocytic intermediates. Actin dynamics and actin-binding proteins are involved in this process, although their detailed role remains controversial. We show here that a dynamic, branched actin network is sufficient, in absence of any membrane-associated proteins, to initiate both inward and outward membrane deformation. With actin polymerization triggered at the membrane of liposomes, we produce inward filopodia-like structures at low tension, while outward endocytosis-like structures are robustly generated regardless of tension. Our results are reminiscent of endocytosis in mammalian cells, where actin polymerization forces are required when membrane tension is increased, and in yeast, where they are always required to overcome the opposing turgor pressure. By combining experimental observations with physical modeling, we propose a mechanism for actin-driven endocytosis under high tension or high pressure conditions.

biophysics↗