M8R tropomyosin mutation disrupts actin binding and filament regulation: The beginning affects the middle and end

Dilated cardiomyopathy (DCM) is associated with mutations in cardiomyocyte sarcomeric proteins, including α-tropomyosin. In conjunction with troponin, tropomyosin shifts to regulate actomyosin interactions. Tropomyosin molecules overlap via tropomyosin–tropomyosin head-to-tail associations, forming a continuous strand along the thin filament. These associations are critical for propagation of tropomyosin''s reconfiguration along the thin filament and key for the cooperative switching between heart muscle contraction and relaxation. Here, we tested perturbations in tropomyosin structure, biochemistry, and function caused by the DCM-linked mutation, M8R, which is located at the overlap junction. Localized and nonlocalized structural effects of the mutation were found in tropomyosin that ultimately perturb its thin filament regulatory function. Comparison of mutant and WT α-tropomyosin was carried out using in vitro motility assays, CD, actin co-sedimentation, and molecular dynamics simulations. Regulated thin filament velocity measurements showed that the presence of M8R tropomyosin decreased calcium sensitivity and thin filament cooperativity. The co-sedimentation of actin and tropomyosin showed weakening of actin-mutant tropomyosin binding. The binding of troponin T''s N terminus to the actin-mutant tropomyosin complex was also weakened. CD and molecular dynamics indicate that the M8R mutation disrupts the four-helix bundle at the head-to-tail junction, leading to weaker tropomyosin–tropomyosin binding and weaker tropomyosin–actin binding. Molecular dynamics revealed that altered end-to-end bond formation has effects extending toward the central region of the tropomyosin molecule, which alter the azimuthal position of tropomyosin, likely disrupting the mutant thin filament response to calcium. These results demonstrate that mutation-induced alterations in tropomyosin–thin filament interactions underlie the altered regulatory phenotype and ultimately the pathogenesis of DCM.     

Integrating multiple data sources to assess the distribution and abundance of bottlenose dolphins Tursiops truncatus in Scottish waters

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Demographic response of tundra small mammals to a snow fencing experiment

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A layover stop in the African American great migration: identity,ruination, and memory

Gone Home challenges dominant representations of Appalachia as white, placing in the foreground the life histories of African Americans who hail from the coalfields of eastern Kentucky. The book reveals how mass migration shapes subjectivities, examining two generations of African Americans who migrated into and out of the coalfields between 1910 and 1970. The coalfields were a layover stop in an intergenerational migration from the rural South to urban centres in the North, Midwest, and West. This review reflects on the significance of the layover stop, both empirically and conceptually. One of the key contributions of Gone Home is to show how the layover stop was important not only in terms of time and place, but also in terms of collective identity. The review concludes by reflecting on interconnected themes of industrial ruination and intergenerational memory.     

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Rhamnogalacturonan II structure shows variation in the side chains monosaccharide composition and methylation status within and across different plant species

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