Tropomyosin (Tpm) isoforms are the master regulators of the functions of individual actin filaments in fungi and metazoans. Tpms are coiled-coil parallel dimers that form a head-to-tail polymer along ...the length of actin filaments. Yeast only has two Tpm isoforms, whereas mammals have over 40. Each cytoskeletal actin filament contains a homopolymer of Tpm homodimers, resulting in a filament of uniform Tpm composition along its length. Evidence for this 'master regulator' role is based on four core sets of observation. First, spatially and functionally distinct actin filaments contain different Tpm isoforms, and recent data suggest that members of the formin family of actin filament nucleators can specify which Tpm isoform is added to the growing actin filament. Second, Tpms regulate whole-organism physiology in terms of morphogenesis, cell proliferation, vesicle trafficking, biomechanics, glucose metabolism and organ size in an isoform-specific manner. Third, Tpms achieve these functional outputs by regulating the interaction of actin filaments with myosin motors and actin-binding proteins in an isoform-specific manner. Last, the assembly of complex structures, such as stress fibers and podosomes involves the collaboration of multiple types of actin filament specified by their Tpm composition. This allows the cell to specify actin filament function in time and space by simply specifying their Tpm isoform composition.
The physiological function of actin filaments is challenging to dissect because of the pleiotropic impact of global disruption of the actin cytoskeleton. Tropomyosin isoforms have provided a unique ...opportunity to address this issue. A substantial fraction of actin filaments in animal cells consist of co-polymers of actin with specific tropomyosin isoforms which determine the functional capacity of the filament. Genetic manipulation of the tropomyosins has revealed isoform specific roles and identified the physiological function of the different actin filament types based on their tropomyosin isoform composition. Surprisingly, there is remarkably little redundancy between the tropomyosins resulting in highly penetrant impacts of both ectopic overexpression and knockout of isoforms. The physiological roles of the tropomyosins cover a broad range from development and morphogenesis to cell migration and specialised tissue function and human diseases.
Mechanical force generation plays an essential role in many cellular functions, including mitosis. Actomyosin contractile forces mediate changes in cell shape in mitosis and are implicated in mitotic ...spindle integrity via cortical tension. An unbiased screen of 150 small molecules that impact actin organization and 32 anti-mitotic drugs identified two molecular targets, Rho kinase (ROCK) and tropomyosin 3.1/2 (Tpm3.1/2), whose inhibition has the greatest impact on mitotic cortical tension. The converse was found for compounds that depolymerize microtubules. Tpm3.1/2 forms a co-polymer with mitotic cortical actin filaments, and its inhibition prevents rescue of multipolar spindles induced by anti-microtubule chemotherapeutics. We examined the role of mitotic cortical tension in this rescue mechanism. Inhibition of ROCK and Tpm3.1/2 and knockdown (KD) of cortical nonmuscle myosin 2A (NM2A), all of which reduce cortical tension, inhibited rescue of multipolar mitotic spindles, further implicating cortical tension in the rescue mechanism. GEF-H1 released from microtubules by depolymerization increased cortical tension through the RhoA pathway, and its KD also inhibited rescue of multipolar mitotic spindles. We conclude that microtubule depolymerization by anti-cancer drugs induces cortical-tension-based rescue to ensure integrity of the mitotic bipolar spindle mediated via the RhoA pathway. Central to this mechanism is the dependence of NM2A on Tpm3.1/2 to produce the functional engagement of actin filaments responsible for cortical tension.
•Rho kinase and tropomyosin Tpm3.1 are key determinants of mitotic cortical tension•Cortical tension mediates rescue of the bipolar spindle from microtubule asters•Agents that compromise mitotic cortical tension inhibit bipolar spindle rescue•Microtubule depolymerization releases active GEF-H1 to promote cortical tension
Mitotic cortical tension is mediated by Rho kinase, Tpm3.1/actin filaments, nonmuscle myosin 2A, and GEF-H1. Wang et al. demonstrate that agents that compromise any one of these mediators collaborate with microtubule depolymerizers to promote microtubule aster formation and inhibit generation of a bipolar mitotic spindle.
The actin cytoskeleton is the primary polymer system within cells responsible for regulating cellular stiffness. While various actin binding proteins regulate the organization and dynamics of the ...actin cytoskeleton, the proteins responsible for regulating the mechanical properties of cells are still not fully understood. In the present study, we have addressed the significance of the actin associated protein, tropomyosin (Tpm), in influencing the mechanical properties of cells. Tpms belong to a multi-gene family that form a co-polymer with actin filaments and differentially regulate actin filament stability, function and organization. Tpm isoform expression is highly regulated and together with the ability to sort to specific intracellular sites, result in the generation of distinct Tpm isoform-containing actin filament populations. Nanomechanical measurements conducted with an Atomic Force Microscope using indentation in Peak Force Tapping in indentation/ramping mode, demonstrated that Tpm impacts on cell stiffness and the observed effect occurred in a Tpm isoform-specific manner. Quantitative analysis of the cellular filamentous actin (F-actin) pool conducted both biochemically and with the use of a linear detection algorithm to evaluate actin structures revealed that an altered F-actin pool does not absolutely predict changes in cell stiffness. Inhibition of non-muscle myosin II revealed that intracellular tension generated by myosin II is required for the observed increase in cell stiffness. Lastly, we show that the observed increase in cell stiffness is partially recapitulated in vivo as detected in epididymal fat pads isolated from a Tpm3.1 transgenic mouse line. Together these data are consistent with a role for Tpm in regulating cell stiffness via the generation of specific populations of Tpm isoform-containing actin filaments.
The actin cytoskeleton is the primary driver of cellular adhesion and mechanosensing due to its ability to generate force and sense the stiffness of the environment. At the cell's leading edge, ...severing of the protruding Arp2/3 actin network generates a specific actin/tropomyosin (Tpm) filament population that controls lamellipodial persistence. The interaction between these filaments and adhesion to the environment is unknown. Using cellular cryo-electron tomography we resolve the ultrastructure of the Tpm/actin copolymers and show that they specifically anchor to nascent adhesions and are essential for focal adhesion assembly. Re-expression of Tpm1.8/1.9 in transformed and cancer cells is sufficient to restore cell-substrate adhesions. We demonstrate that knock-out of Tpm1.8/1.9 disrupts the formation of dorsal actin bundles, hindering the recruitment of α-actinin and non-muscle myosin IIa, critical mechanosensors. This loss causes a force-generation and proliferation defect that is notably reversed when cells are grown on soft surfaces. We conclude that Tpm1.8/1.9 suppress the metastatic phenotype, which may explain why transformed cells naturally downregulate this Tpm subset during malignant transformation.
Actin filament functional diversity is paralleled by variation in the composition of isoforms of tropomyosin in these filaments. Although the role of tropomyosin is well understood in skeletal ...muscle, where it regulates the actin–myosin interaction, its role in the cytoskeleton has been obscure. The intracellular sorting of tropomyosin isoforms indicated a role in spatial specialization of actin filament function. Genetic manipulation and protein chemistry studies have confirmed that these isoforms are functionally distinct. Tropomyosins differ in their recruitment of myosin motors and their interaction with actin filament regulators such as ADF-cofilin. Tropomyosin isoforms have therefore provided a powerful mechanism to diversify actin filament function in different intracellular compartments.
Tropomyosins Gunning, Peter W.; Hardeman, Edna C.
CB/Current biology,
01/2017, Letnik:
27, Številka:
1
Journal Article
Recenzirano
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The actin cytoskeleton provides not only the underpinning for cell architecture but also mechanical force and the ability to drive movement of cells and their organelles. It is tempting to think of ...it simply as a set of stable structural elements, but nothing could be further from the truth. The cells of our bodies are continually remodelling their architecture by responding to a range of imposed biomechanical forces and intracellular functional demands. Studies of the dynamic and functional properties of the actin cytoskeleton have been dominated by a focus on actin and the view that actin filaments are essentially ‘generic’. However, the ‘other’ component of most actin filaments in animals — tropomyosin — is coming into prominence. With this discovery is the realisation that far from being generic, actin filaments have their own functional individuality provided to them by their associated tropomyosin. This is changing the way we understand and study the actin cytoskeleton and has delivered a new therapeutic opportunity in what had come to be considered a ‘no-go zone’.
Here, Hardeman and Gunning discuss the emerging view that tropomyosins specify the functional properties of individual cytoskeletal actin filaments in an isoform-specific manner.
Cancer cells have now been shown to lack rigidity-sensing due to alteration in cytoskeletal sensor proteins, but can be reversed from a transformed to a rigidity-dependent growth state by the sensor ...proteins, resulting in restoration of contractility and adhesion.
The negative impact of irradiation or diet on the metabolic and immune profiles of cancer survivors have been previously demonstrated. The gut microbiota plays a critical role in regulating these ...functions and is highly sensitive to cancer therapies. The aim of this study was to investigate the effect of irradiation and diet on the gut microbiota and metabolic or immune functions. We exposed C57Bl/6J mice to a single dose of 6 Gy radiation and after 5 weeks, fed them a chow or high-fat diet (HFD) for 12 weeks. We characterised their faecal microbiota, metabolic (whole body and adipose tissue) functions, and systemic (multiplex cytokine, chemokine assay, and immune cell profiling) and adipose tissue inflammatory profiles (immune cell profiling). At the end of the study, we observed a compounding effect of irradiation and diet on the metabolic and immune profiles of adipose tissue, with exposed mice fed a HFD displaying a greater inflammatory signature and impaired metabolism. Mice fed a HFD also showed altered microbiota, irrespective of irradiation status. An altered diet may exacerbate the detrimental effects of irradiation on both the metabolic and inflammatory profiles. This could have implications for the diagnosis and prevention of metabolic complications in cancer survivors exposed to radiation.
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