Reconstitution of cytolinker-mediated crosstalk between actin and vimentin

European Journal of Cell Biology, Год журнала: 2024, Номер 103(2), С. 151403 - 151403

Опубликована: Март 12, 2024

Cell shape and motility are determined by the cytoskeleton, an interpenetrating network of actin filaments, microtubules, intermediate filaments. The biophysical properties each filament type individually have been studied extensively cell-free reconstitution. By contrast, interactions between three cytoskeletal networks relatively unexplored. They coupled via crosslinkers plakin family such as plectin. These challenging proteins for reconstitution because their giant size multidomain structure. Here we engineer a recombinant actin-vimentin crosslinker protein called 'ACTIF' that provides minimal model system plectin, recapitulating its modular design with actin-binding filament-binding domains separated coiled-coil linker dimerisation. We show fluorescence electron microscopy ACTIF has high binding affinity vimentin creates mixed bundles. Rheology measurements ACTIF-mediated crosslinking strongly stiffens composites. Finally, demonstrate modularity this approach creating variant domain Adenomatous Polyposis Coli. Our engineering new characterization understanding mechanical synergy in mesenchymal cells.

Язык: Английский

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Structural determinants of intermediate filament mechanics

Current Opinion in Cell Biology, Год журнала: 2024, Номер 89, С. 102375 - 102375

Опубликована: Июнь 7, 2024

Intermediate filaments (IFs) are integral to the cell cytoskeleton, supporting cellular mechanical stability. Unlike other cytoskeletal components, detailed structure of assembled IFs has yet be resolved. This review highlights new insights, linking complex IF hierarchical assembly their properties and impact on functions. While we focus vimentin IFs, draw comparisons keratins, showcasing distinctive structural features that underlie unique responses.

Язык: Английский

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Heterogeneous force response of chromatin in isolated nuclei

Cell Reports, Год журнала: 2024, Номер 43(10), С. 114852 - 114852

Опубликована: Окт. 1, 2024

A quantitative description of nuclear mechanics is crucial for understanding its role in force sensing within eukaryotic cells. Recent studies indicate that the chromatin nucleus cannot be treated as a homogeneous material. To elucidate material properties, we combine optical tweezers manipulation isolated nuclei with multi-color fluorescence imaging lamin and to map response local deformations. Force spectroscopy reveals strain stiffening an exponential dependence, well described by hierarchical chain model. Simultaneously, data show higher compliance compared envelope at strains <30%. Micrococcal nuclease (MNase) digestion results softening can captured our Additionally, observe stretching responses showing lipid tether signature, suggesting these tethers originate from membrane. Our combined approach allows us while mapping deformation lamin, (eu)chromatin,

Язык: Английский

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The redox-responsive roles of intermediate filaments in cellular stress detection, integration and mitigation

Current Opinion in Cell Biology, Год журнала: 2023, Номер 86, С. 102283 - 102283

Опубликована: Ноя. 20, 2023

Intermediate filaments are critical for cell and tissue homeostasis stress responses. Cytoplasmic intermediate form versatile dynamic assemblies that interconnect cellular organelles, participate in signaling protect cells tissues against stress. Here we have focused on their involvement redox oxidative stress, which arises numerous pathophysiological situations. We pay special attention to type III filaments, mainly vimentin, because it provides a physical interface signaling, responses mechanosensing. Vimentin possesses single cysteine residue is target multiple oxidants electrophiles. This conserved fine tunes vimentin assembly, response crosstalk with other structures. integrate evidence from the filament biology fields propose as sentinel networks of cell. To support this, appraise how detects orchestrates electrophilic

Язык: Английский

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Extracellular vimentin as a modulator of the immune response and an important player during infectious diseases

Immunology and Cell Biology, Год журнала: 2024, Номер 102(3), С. 167 - 178

Опубликована: Янв. 11, 2024

Abstract Vimentin, an intermediate filament protein primarily recognized for its intracellular role in maintaining cellular structure, has recently garnered increased attention and emerged as a pivotal extracellular player immune regulation host–pathogen interactions. While the functions of vimentin were initially overshadowed by cytoskeletal role, accumulating evidence now highlights significance diverse physiological pathological events. This review explores multifaceted modulating responses orchestrating interactions between host cells pathogens. It delves into mechanisms underlying vimentin's release milieu, elucidating unconventional secretion pathways identifying critical molecular triggers. In addition, future perspectives using diagnostics target treatment diseases are discussed.

Язык: Английский

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How cytoskeletal crosstalk makes cells move: Bridging cell-free and cell studies

Biophysics Reviews, Год журнала: 2024, Номер 5(2)

Опубликована: Июнь 1, 2024

Cell migration is a fundamental process for life and highly dependent on the dynamical mechanical properties of cytoskeleton. Intensive physical biochemical crosstalk among actin, microtubules, intermediate filaments ensures their coordination to facilitate enable migration. In this review, we discuss different aspects that govern cell provide, each aspect, novel perspective by juxtaposing two complementary approaches biophysical study cytoskeletal crosstalk: live-cell studies (often referred as top-down studies) cell-free bottom-up studies). We summarize main findings from both experimental approaches, provide our bridging perspectives address open questions how governs makes cells move.

Язык: Английский

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Vimentin promotes collective cell migration through collagen networks via increased matrix remodeling and spheroid fluidity

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Июнь 17, 2024

The intermediate filament (IF) protein vimentin is associated with many diseases phenotypes of enhanced cellular migration and aggressive invasion through the extracellular matrix (ECM) tissues, but vimentin's role in in-vivo cell still largely unclear. Vimentin important for proper adhesion force generation, which are critical to migration; yet cytoskeleton also hinders ability cells squeeze small pores ECM, resisting migration. To identify collective migration, we generate spheroids wide-type vimentin-null mouse embryonic fibroblasts (mEFs) embed them a 3D collagen matrix. We find that loss significantly impairs spheroid collectively expand networks remodel network. Traction analysis reveals null exert less contractile than their wild-type counterparts. In addition, made mEFs only unit length filaments (ULFs) exhibit similar behavior as spheroids, suggesting filamentous required promote vimentin-mediated expansion dependent on metalloproteinase (MMP) degradation Further, vertex model simulation embedded ECM indicates behave more fluid-like, enabling active pulling reconstructing surrounding Altogether, these results signify VIF plays enhancing migratory persistence environments MMP transportation tissue fluidity.

Язык: Английский

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Reorganizing chromatin by cellular deformation

Current Opinion in Cell Biology, Год журнала: 2024, Номер 90, С. 102408 - 102408

Опубликована: Авг. 8, 2024

Язык: Английский

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Intermediate filaments at a glance

Journal of Cell Science, Год журнала: 2024, Номер 137(16)

Опубликована: Авг. 15, 2024

Intermediate filaments (IFs) comprise a large family of versatile cytoskeletal proteins, divided into six subtypes with tissue-specific expression patterns. IFs have wide repertoire cellular functions, including providing structural support to cells, as well active roles in mechanical and signaling pathways. Consequently, defects are associated more than 100 diseases. In this Cell Science at Glance article, we discuss the established classes their general features, functions beyond support, recent advances field. We also highlight involvement disease potential use clinical markers pathological conditions. Finally, provide our view on current knowledge gaps future directions IF

Язык: Английский

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Actin and vimentin jointly control cell viscoelasticity and compression stiffening

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2025, Номер unknown

Опубликована: Янв. 2, 2025

Abstract The mechanical properties of cells are governed by the cytoskeleton, a dynamic network actin filaments, intermediate and microtubules. Understanding individual collective contributions these three different cytoskeletal elements is essential to elucidate how maintain integrity during deformation. Here we use custom single-cell rheometer identify distinct vimentin viscoelastic nonlinear elastic response uniaxial compression. We used mouse embryonic fibroblasts (MEFs) isolated from wild type (WT) knockout (vim -/-) mice in combination with chemical treatments manipulate polymerization contractility. show through small amplitude oscillatory measurements strain ramp tests that vimentin, often overlooked cellular mechanics, plays role comparable maintaining cell stiffness resisting large compressive forces. However, appears be more important than determining energy dissipation. Finally comparing enucleated compression stiffening originates while nucleus play little this. Our findings provide insight into networks collectively determine cells, providing basis understand cytoskeleton ability resist external as well internal Significance statement A cell’s stress largely networks, but their relative contribution viscoelasticity deformations poorly characterized. reveal have an almost equal strain-stiffen under This work underscores cytoskeleton’s central mechanics synergy between framework for understanding components coordinate structural respond environments.

Язык: Английский

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