Quantitation of the distribution and flux of myosin-II during cytokinesis

BACKGROUND: During cytokinesis, the cell's equator contracts against the cell's global stiffness. Identifying the biochemical basis for these mechanical parameters is essential for understanding how cells divide. To achieve this goal, the distribution and flux of the cell division machinery must be quantified. Here we report the first quantitative analysis of the distribution and flux of myosin-II, an essential element of the contractile ring. RESULTS: The fluxes of myosin-II in the furrow cortex, the polar cortex, and the cytoplasm were examined using ratio imaging of GFP fusion proteins expressed in Dictyostelium. The peak concentration of GFP-myosin-II in the furrow cortex is 1.8-fold higher than in the polar cortex and 2.0-fold higher than in the cytoplasm. The myosin-II in the furrow cortex, however, represents only 10% of the total cellular myosin-II. An estimate of the minimal amount of this motor needed to produce the required force for cell cleavage fits well with this 10% value. The cell may, therefore, regulate the amount of myosin-II sent to the furrow cortex in accordance with the amount needed there. Quantitation of the distribution and flux of a mutant myosin-II that is defective in phosphorylation-dependent thick filament disassembly confirms that heavy chain phosphorylation regulates normal recruitment to the furrow cortex. CONCLUSION: The analysis indicates that myosin-II flux through the cleavage furrow cortex is regulated by thick filament phosphorylation. Further, the amount of myosin-II observed in the furrow cortex is in close agreement with the amount predicted to be required from a simple theoretical analysis

A Concept for Robust, High Density Terminal Air Traffic Operations

This paper describes a concept for future high-density, terminal air traffic operations that has been developed by interpreting the Joint Planning and Development Office s vision for the Next Generation (NextGen) Air Transportation System and coupling it with emergent NASA and other technologies and procedures during the NextGen timeframe. The concept described in this paper includes five core capabilities: 1) Extended Terminal Area Routing, 2) Precision Scheduling Along Routes, 3) Merging and Spacing, 4) Tactical Separation, and 5) Off-Nominal Recovery. Gradual changes are introduced to the National Airspace System (NAS) by phased enhancements to the core capabilities in the form of increased levels of automation and decision support as well as targeted task delegation. NASA will be evaluating these conceptual technological enhancements in a series of human-in-the-loop simulations and will accelerate development of the most promising capabilities in cooperation with the FAA through the Efficient Flows Into Congested Airspace Research Transition Team

Automated characterization of cell shape changes during amoeboid motility by skeletonization

<p>Abstract</p> <p>Background</p> <p>The ability of a cell to change shape is crucial for the proper function of many cellular processes, including cell migration. One type of cell migration, referred to as amoeboid motility, involves alternating cycles of morphological expansion and retraction. Traditionally, this process has been characterized by a number of parameters providing global information about shape changes, which are insufficient to distinguish phenotypes based on local pseudopodial activities that typify amoeboid motility.</p> <p>Results</p> <p>We developed a method that automatically detects and characterizes pseudopodial behavior of cells. The method uses skeletonization, a technique from morphological image processing to reduce a shape into a series of connected lines. It involves a series of automatic algorithms including image segmentation, boundary smoothing, skeletonization and branch pruning, and takes into account the cell shape changes between successive frames to detect protrusion and retraction activities. In addition, the activities are clustered into different groups, each representing the protruding and retracting history of an individual pseudopod.</p> <p>Conclusions</p> <p>We illustrate the algorithms on movies of chemotaxing <it>Dictyostelium </it>cells and show that our method makes it possible to capture the spatial and temporal dynamics as well as the stochastic features of the pseudopodial behavior. Thus, the method provides a powerful tool for investigating amoeboid motility.</p

Understanding the Cooperative Interaction between Myosin II and Actin Cross-Linkers Mediated by Actin Filaments during Mechanosensation

AbstractMyosin II is a central mechanoenzyme in a wide range of cellular morphogenic processes. Its cellular localization is dependent not only on signal transduction pathways, but also on mechanical stress. We suggest that this stress-dependent distribution is the result of both the force-dependent binding to actin filaments and cooperative interactions between bound myosin heads. By assuming that the binding of myosin heads induces and/or stabilizes local conformational changes in the actin filaments that enhances myosin II binding locally, we successfully simulate the cooperative binding of myosin to actin observed experimentally. In addition, we can interpret the cooperative interactions between myosin and actin cross-linking proteins observed in cellular mechanosensation, provided that a similar mechanism operates among different proteins. Finally, we present a model that couples cooperative interactions to the assembly dynamics of myosin bipolar thick filaments and that accounts for the transient behaviors of the myosin II accumulation during mechanosensation. This mechanism is likely to be general for a range of myosin II-dependent cellular mechanosensory processes

Dynacortin facilitates polarization of chemotaxing cells

<p>Abstract</p> <p>Background</p> <p>Cell shape changes during cytokinesis and chemotaxis require regulation of the actin cytoskeletal network. Dynacortin, an actin cross-linking protein, localizes to the cell cortex and contributes to cortical resistance, thereby helping to define the cell shape changes of cytokinesis. Dynacortin also becomes highly enriched in cortical protrusions, which are sites of new actin assembly.</p> <p>Results</p> <p>We studied the effect of dynacortin on cell motility during chemotaxis and on actin dynamics <it>in vivo </it>and <it>in vitro</it>. Dynacortin enriches with the actin, particularly at the leading edge of chemotaxing cells. Cells devoid of dynacortin do not become as polarized as wild-type control cells but move with similar velocities as wild-type cells. In particular, they send out multiple pseudopods that radiate at a broader distribution of angles relative to the chemoattractant gradient. Wild-type cells typically only send out one pseudopod at a time that does not diverge much from 0° on average relative to the gradient. Though <it>dynacortin</it>-deficient cells show normal bulk (whole-cell) actin assembly upon chemoattractant stimulation, dynacortin can promote actin assembly <it>in vitro</it>. By fluorescence spectroscopy, co-sedimentation and transmission electron microscopy, dynacortin acts as an actin scaffolder in which it assembles actin monomers into polymers with a stoichiometry of 1 Dyn₂:1 actin under salt conditions that disfavor polymer assembly.</p> <p>Conclusion</p> <p>Dynacortin contributes to cell polarization during chemotaxis. By cross-linking and possibly stabilizing actin polymers, dynacortin also contributes to cortical viscoelasticity, which may be critical for establishing cell polarity. Though not essential for directional sensing or motility, dynacortin is required to establish cell polarity, the third core feature of chemotaxis.</p

Mechanoaccumulative elements of the mammalian actin cytoskeleton

To change shape, divide, form junctions, and migrate, cells reorganize their cytoskeletons in response to changing mechanical environments [1-4]. Actin cytoskeletal elements, including myosin II motors and actin crosslinkers, structurally remodel and activate signaling pathways in response to imposed stresses [5-9]. Recent studies demonstrate the importance of force-dependent structural rearrangement of α-catenin in adherens junctions [10] and vinculin's molecular clutch mechanism in focal adhesions [11]. However, the complete landscape of cytoskeletal mechanoresponsive proteins and the mechanisms by which these elements sense and respond to force remain to be elucidated. To find mechanosensitive elements in mammalian cells, we examined protein relocalization in response to controlled external stresses applied to individual cells. Here, we show that non-muscle myosin II, α-actinin, and filamin accumulate to mechanically stressed regions in cells from diverse lineages. Using reaction-diffusion models for force-sensitive binding, we successfully predicted which mammalian α-actinin and filamin paralogs would be mechanoaccumulative. Furthermore, a Goldilocks zone must exist for each protein where the actin-binding affinity must be optimal for accumulation. In addition, we leveraged genetic mutants to gain a molecular understanding of the mechanisms of α-actinin and filamin catch-bonding behavior. Two distinct modes of mechanoaccumulation can be observed: a fast, diffusion-based accumulation and a slower, myosin II-dependent cortical flow phase that acts on proteins with specific binding lifetimes. Finally, we uncovered cell-type and cell-cycle-stage-specific control of the mechanosensation of myosin IIB, but not myosin IIA or IIC. Overall, these mechanoaccumulative mechanisms drive the cell's response to physical perturbation during proper tissue development and disease

Choral Ensembles Spring Concert

This Kennesaw State University School of Music performance features Chamber Singers, Men\u27s Ensemble, and University Chorale directed by Dr. Leslie Blackwell, Director of Choral Activities and Professor of Music and Music Education.https://digitalcommons.kennesaw.edu/musicprograms/2054/thumbnail.jp

Penrose Limits and Non-local theories

We investigate Penrose limits of two classes of non-local theories, little string theories and non-commutative gauge theories. Penrose limits of the near-horizon geometry of NS5-branes help to shed some light on the high energy spectrum of little string theories. We attempt to understand renormalization group flow in these theories by considering Penrose limits wherein the null geodesic also has a radial component. In particular, we demonstrate that it is possible to construct a pp-wave spacetime which interpolates between the linear dilaton and the AdS regions for the Type IIA NS5-brane. Similar analysis is considered for the holographic dual geometry to non-commutative field theories.Comment: 27 pages, LaTeX; v2: added reference