bioRxiv Science⌕ Search

Biology subjects

Henon, S.

Publications and source records attributed to Henon, S..

4 recordsLinked to original sources

Nuclear pore complex distribution on the nuclear envelope: insights into curvature, chromatin, and actin contributions

The Nuclear Pore Complex (NPC) is an ancestral feature of eukaryotic cells, essential for the exchange of material and information between cytoplasm and nucleoplasm. While, typical eukaryotic cells harbor thousands of NPCs covering the nucleus surface, a quantitative characterization of their spatial distribution remains lacking. Taking advantage of super-resolution microscopy, we imaged and segmented individual NPCs in myoblast cells to reconstruct the nuclear surface. Our spatial statistics analysis of NPCs on the nuclear surface reveals the existence of a 400nm repulsion length between NPCs, likely reflecting the mesh size of the underlying lamin network. Furthermore, we identify three key correlates of NPC density. (1) Myoblast actin fibres, which can indent the nucleus, correlate with a decrease in NPC surface density, contrary to what was reported in previous studies; (2) Regions of the nuclear envelope near large heterochromatin domains exhibit reduced NPC surface density; and (3) There is a non-monotonous relationship between NPC density and the nuclear envelope curvature. This last observation is consistent with the existence of NPC-preferred curvature suggesting curvature sensing properties. In conclusion, this study provides new insights into the spatial distribution of NPCs, shedding light on possible new determinants of NPC spatial distribution.

biophysics↗

Multiparameters dependance of tissue shape maintenance in myoblasts multicellular aggregates: the role of intermediate filaments.

Liquid and elastic behavior of tissues drives their morphology and their response to the environment. They appear as the first insight on tissue mechanics. We explore the role of individual cell properties on spheroids of mouse muscle precursor cells by developing a fully automated surface tension and Youngs modulus measurement system. Flattening multicellular aggregates under magnetic constraint, we show that rigidity and surface tension act as highly sensitive macroscopic reporters closely related to microscopic local tension and effective adhesion. Shedding light on the major contributions of acto-myosin contractility, actin organization and intercellular adhesions, we reveal the role of desmin organization on the macroscopic mechanics of this tissue model.

biophysics↗

The impact of cell geometry and the cytoskeleton on the nucleo-cytoplasmic localisation of the SMYD3 methyltransferase suggests that the epigenetic machinery is mechanosensitive

Mechanical cues from the cellular microenvironment are converted into biochemical signals controlling diverse cell behaviours, including growth and differentiation. But it is still unclear how mechanotransduction ultimately affects nuclear readouts, genome function and transcriptional programs. Key signaling pathways and transcription factors can be activated, and can relocalize to the nucleus, upon mechanosensing. Here, we tested the hypothesis that epigenetic regulators, such as methyltransferase enzymes, might also contribute to mechanotransduction. We found that the SMYD3 lysine methyltransferase is spatially redistributed dependent on cell geometry (cell shape and aspect ratio) in murine myoblasts. Specifically, elongated rectangles were less permissive than square shapes to SMYD3 nuclear accumulation, via reduced nuclear import. Notably, SMYD3 has both nuclear and cytoplasmic substrates. The distribution of SMYD3 in response to cell geometry correlated with cytoplasmic and nuclear lysine tri-methylation (Kme3) levels, but not Kme2. Moreover, drugs targeting cytoskeletal acto-myosin induced nuclear accumulation of Smyd3. We also observed that square vs rectangular geometry impacted the nuclear-cytoplasmic relocalisation of several mechano-sensitive proteins, notably YAP/TAZ proteins and the SETDB1 methyltransferase. Thus, mechanical cues from cellular geometric shapes are transduced by a combination of transcription factors and epigenetic regulators shuttling between the cell nucleus and cytoplasm.

biophysics↗

The nature and intensity of mechanical stimulation drive different dynamics of MRTF-A nuclear redistribution after actin remodeling in myoblasts

Serum response factor and its cofactor myocardin-related transcription factor (MRTF) are key elements of muscle-mass adaptation to workload. The transcription of target genes is activated when MRTF is present in the nucleus. The localization of MRTF is controlled by its binding to G-actin. Thus, the pathway can be mechanically activated through the mechanosensitivity of the actin cytoskeleton. The pathway has been widely investigated from a biochemical point of view, but its mechanical activation and the timescales involved are poorly understood. Here, we applied local and global mechanical cues to myoblasts through two custom-built set-ups, magnetic tweezers and stretchable substrates. Both induced nuclear accumulation of MRTF-A. However, the dynamics of the response varied with the nature and level of mechanical stimulation and correlated with the polymerization of different actin sub-structures. Local repeated force induced local actin polymerization and nuclear accumulation of MRTF-A by 30 minutes, whereas a global static strain induced both rapid (minutes) transient nuclear accumulation, associated with the polymerization of an actin cap above the nucleus, and long-term (hours) accumulation, with a global increase in polymerized actin. Conversely, high strain induced actin depolymerization at intermediate times, associated with cytoplasmic MRTF accumulation.

biophysics↗