scholarly journals Effects of dynein on microtubule mechanics and centrosome positioning

2011 ◽  
Vol 22 (24) ◽  
pp. 4834-4841 ◽  
Author(s):  
Jun Wu ◽  
Gaurav Misra ◽  
Robert J. Russell ◽  
Anthony J. C. Ladd ◽  
Tanmay P. Lele ◽  
...  
Keyword(s):  
Force Balance ◽  
Dynamic Force ◽  
Cell Periphery ◽  
Friction Forces ◽  
Dynein Motor ◽  
Elastic Forces ◽  
Pulling Forces ◽  
Capillary Endothelial Cells ◽  
Pulling Force ◽  
Shape Changes

To determine forces on intracellular microtubules, we measured shape changes of individual microtubules following laser severing in bovine capillary endothelial cells. Surprisingly, regions near newly created minus ends increased in curvature following severing, whereas regions near new microtubule plus ends depolymerized without any observable change in shape. With dynein inhibited, regions near severed minus ends straightened rapidly following severing. These observations suggest that dynein exerts a pulling force on the microtubule that buckles the newly created minus end. Moreover, the lack of any observable straightening suggests that dynein prevents lateral motion of microtubules. To explain these results, we developed a model for intracellular microtubule mechanics that predicts the enhanced buckling at the minus end of a severed microtubule. Our results show that microtubule shapes reflect a dynamic force balance in which dynein motor and friction forces dominate elastic forces arising from bending moments. A centrosomal array of microtubules subjected to dynein pulling forces and resisted by dynein friction is predicted to center on the experimentally observed time scale, with or without the pushing forces derived from microtubule buckling at the cell periphery.

scholarly journals Cortical dynein pulling mechanism is regulated by differentially targeted attachment molecule Num1

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Safia Omer ◽  
Samuel R Greenberg ◽  
Wei-Lih Lee
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Spatial Distribution ◽  
Direct Evidence ◽  
Critical Factor ◽  
Spindle Positioning ◽  
Dynein Motor ◽  
Independent Population ◽  
Bud Neck ◽  
Pulling Forces ◽  
Pulling Force

Cortical dynein generates pulling forces via microtubule (MT) end capture-shrinkage and lateral MT sliding mechanisms. In Saccharomyces cerevisiae, the dynein attachment molecule Num1 interacts with endoplasmic reticulum (ER) and mitochondria to facilitate spindle positioning across the mother-bud neck, but direct evidence for how these cortical contacts regulate dynein-dependent pulling forces is lacking. We show that loss of Scs2/Scs22, ER tethering proteins, resulted in defective Num1 distribution and loss of dynein-dependent MT sliding, the hallmark of dynein function. Cells lacking Scs2/Scs22 performed spindle positioning via MT end capture-shrinkage mechanism, requiring dynein anchorage to an ER- and mitochondria-independent population of Num1, dynein motor activity, and CAP-Gly domain of dynactin Nip100/p150Glued subunit. Additionally, a CAAX-targeted Num1 rescued loss of lateral patches and MT sliding in the absence of Scs2/Scs22. These results reveal distinct populations of Num1 and underline the importance of their spatial distribution as a critical factor for regulating dynein pulling force.

Analytical Modelling of the Dynamics of the Four-Bar Mechanism: A Comparison of Some Methods

2018 ◽  
Author(s):  
J. P. Meijaard ◽  
V. van der Wijk
Keyword(s):  
Virtual Work ◽  
Equations Of Motion ◽  
Differential Algebraic Equations ◽  
Force Balance ◽  
Dynamic Force ◽  
Algebraic Equations ◽  
Principle Of Virtual Work ◽  
Principal Vector ◽  
Reaction Forces ◽  
Principal Points

Some thoughts about different ways of formulating the equations of motion of a four-bar mechanism are communicated. Four analytic methods to derive the equations of motion are compared. In the first method, Lagrange’s equations in the traditional form are used, and in a second method, the principle of virtual work is used, which leads to equivalent equations. In the third method, the loop is opened, principal points and a principal vector linkage are introduced, and the equations are formulated in terms of these principal vectors, which leads, with the introduced reaction forces, to a system of differential-algebraic equations. In the fourth method, equivalent masses are introduced, which leads to a simpler system of principal points and principal vectors. By considering the links as pseudorigid bodies that can have a uniform planar dilatation, a compact form of the equations of motion is obtained. The conditions for dynamic force balance become almost trivial. Also the equations for the resulting reaction moment are considered for all four methods.

scholarly journals Stable tug-of-war between kinesin-1 and cytoplasmic dynein upon different ATP and roadblock concentrations

2020 ◽  
Author(s):  
Gina A. Monzon ◽  
Lara Scharrel ◽  
Ashwin DSouza ◽  
Ludger Santen ◽  
Stefan Diez
Keyword(s):  
Cytoplasmic Dynein ◽  
Gliding Motility ◽  
Force Balance ◽  
Stochastic Simulations ◽  
Transport Systems ◽  
Organelle Transport ◽  
Atp Concentration ◽  
Dynein Motor ◽  
Bidirectional Movement

ABSTRACTThe maintenance of intracellular processes like organelle transport and cell division depend on bidirectional movement along microtubules. These processes typically require kinesin and dynein motor proteins which move with opposite directionality. Because both types of motors are often simultaneously bound to the cargo, regulatory mechanisms are required to ensure controlled directional transport. Recently, it has been shown that parameters like mechanical motor activation, ATP concentration and roadblocks on the microtubule surface differentially influence the activity of kinesin and dynein motors in distinct manners. However, how these parameters affect bidirectional transport systems has not been studied. Here, we investigate the regulatory influence of these three parameter using in vitro gliding motility assays and stochastic simulations. We find that the number of active kinesin and dynein motors determines the transport direction and velocity, but that variations in ATP concentration and roadblock density have no significant effect. Thus, factors influencing the force balance between opposite motors appear to be important, whereas the detailed stepping kinetics and bypassing capabilities of the motors have only little effect.

A Weighted Rearranged Spectral Equation Method for Structural Model Updating

1995 ◽  
Author(s):  
José Roberto F. Arruda ◽  
Carlson Antonio M. Verçosa
Keyword(s):  
Degrees Of Freedom ◽  
Structural Model ◽  
Model Updating ◽  
Structural Connectivity ◽  
Computational Cost ◽  
Force Balance ◽  
Dynamic Force ◽  
Stiffness Coefficients ◽  
Spring System ◽  
Spectral Equation

Abstract A new structural model updating method based on the dynamic force balance is presented. The method consists of rearranging the spectral equation so that measured modes and natural frequencies can be used to compute directly updated stiffness coefficients. The proposed method preserves both the structural connectivity and reciprocity, which translate into sparsity and symmetry of the stiffness matrix, respectively. Large changes in small-valued stiffness coefficients are avoided using parameter weighting in the rearranged spectral equation solution. It is shown that the proposed method produces results which are similar to the results obtained using Alvar Kabe’s method, with the advantages of simpler formulation and smaller computational cost. A simple example of an 8 degrees-of-freedom mass-spring system, originally used by Kabe to present his method, is used here to evaluate the proposed method.

Altering the cellular mechanical force balance results in integrated changes in cell, cytoskeletal and nuclear shape

1992 ◽  
Vol 103 (4) ◽  
pp. 1215-1222 ◽  
Author(s):  
J.R. Sims ◽  
S. Karp ◽  
D.E. Ingber
Keyword(s):  
Shape Control ◽  
Nuclear Shape ◽  
Force Balance ◽  
Dependent Manner ◽  
Integrin Receptors ◽  
Permeabilized Cells ◽  
Capillary Endothelial Cells ◽  
Shape Determination ◽  
Dose Dependent

Studies were carried out with capillary endothelial cells cultured on fibronectin (FN)-coated dishes in order to analyze the mechanism of cell and nuclear shape control by extracellular matrix (ECM). To examine the role of the cytoskeleton in shape determination independent of changes in transmembrane osmotic pressure, membranes of adherent cells were permeabilized with saponin (25 micrograms/ml) using a buffer that maintains the functional integrity of contractile microfilaments. Real-time videomicroscopic studies revealed that addition of 250 microM ATP resulted in time-dependent retraction and rounding of permeabilized cells and nuclei in a manner similar to that observed in intact living cells following detachment using trypsin-EDTA. Computerized image analysis confirmed that permeabilized cells remained essentially rigid in the absence of ATP and that retraction was stimulated in a dose-dependent manner as the concentration of ATP was raised from 10 to 250 microM. Maximal rounding occurred by 30 min with projected cell and nuclear areas being reduced by 69 and 41%, respectively. ATP-induced rounding was also accompanied by a redistribution of microfilaments resulting in formation of a dense net of F-actin surrounding retracted nuclei. Importantly, ATP-stimulated changes in cell, cytoskeletal, and nuclear form were prevented in permeabilized cells using a synthetic myosin peptide (IRICRKG) that has been previously shown to inhibit actomyosin filament sliding in muscle. In contrast, both the rate and extent of cell and nuclear rounding were increased in permeabilized cells exposed to ATP when the soluble FN peptide, GRGDSP, was used to dislodge immobilized FN from cell surface integrin receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Diacylglycerols and PMA induce actin polymerization and distinct shape changes in lymphocytes: relation to fluid pinocytosis and locomotion

1989 ◽  
Vol 93 (3) ◽  
pp. 457-465
Author(s):  
H.U. Keller ◽  
V. Niggli ◽  
A. Zimmermann
Keyword(s):  
Actin Polymerization ◽  
Shape Change ◽  
Second Messengers ◽  
Cell Periphery ◽  
Response Curves ◽  
Blood Lymphocytes ◽  
Different Types ◽  
Net Uptake ◽  
Very High ◽  
Shape Changes

Shape changes have been determined in human blood lymphocytes stimulated with OAG, diC8, PMA, colchicine or the hexapeptide fNLPNTL in short-term assays (30 min). Distinct types of shape-change responses were observed. Colchicine was active in generating a relatively small proportion of polarized lymphocytes (front-tail polarity). OAG, diC8 and PMA produced different types of shape change (non-polar cells with surface projections), and these were closely associated with an increase in actin polymerization and a shift of F-actin into the projections at the cell periphery. The diacylglycerols OAG and diC8 produced biphasic dose-response curves leading to rounding up of cells at very high stimulant concentrations. PMA produced no comparable biphasic response when tested over a much wider concentration range. Though the nonpolar cells with surface projections generated by OAG, diC8 or PMA showed vigorous shape changes, they lacked significant locomotor activity. alpha-Phorbol, 4 alpha-PDD, lumicolchicine or fNLPNTL were inactive. Small blood lymphocytes stimulated by OAG, diC8 or PMA showed a very small increase in the net uptake of FITC-dextran by fluid pinocytosis. Unlike neutrophils, which show a high net uptake, lymphocytes did not concentrate FITC-dextran in large granules, indicating that they do not develop a ‘storage’ compartment in the form of large vesicles. However, small fluorescent spots were consistently found in at least a fraction of blood lymphocytes. The results indicate that stimulated surface movement may be instrumental in fluid pinocytosis. Diacylglycerols may act as second messengers to induce pinocytosis, shape changes and altered actin polymerization in lymphocytes.

scholarly journals Identification of factors influencing insertion characteristics of cochlear implant electrode carriers

2019 ◽  
Vol 5 (1) ◽  
pp. 441-443
Author(s):  
Silke Hügl ◽  
Nina Aldag ◽  
Thomas Lenarz ◽  
Thomas S. Rau ◽  
Alexander Becker ◽  
...  
Keyword(s):  
Cochlear Implant ◽  
Test Bench ◽  
Time Interval ◽  
Dynamic Force ◽  
Friction Forces ◽  
Factors Influencing ◽  
Constant Diameter ◽  
Constant Radius ◽  
Force Components ◽  
Insertion Process

AbstractInsertion studies in artificial cochlea models (aCM) are used for the analysis of insertion characteristics of different cochlear implant electrode carrier (EC) designs by measuring insertion forces. These forces are summed forces due to the measuring position which is directly underneath the aCM. The current hypothesis is that they include dynamic friction forces during the insertion process and the forces needed to bend an initially straight EC into the curved form of the aCM. For the purposes of the present study, straight EC substitutes with a constant diameter of 0.7 mm and 20.5 mm intracochlear length were fabricated out of silicone in two versions with different stiffness by varying the number of embedded wires. The EC substitutes were inserted into three different models made of polytetrafluoroethylene (PTFE), each model showing only one constant radius. Three different insertion speeds were used (0.11 / 0.4 / 1.6 mm/s) with an automated insertion test bench. For each parameter combination (curvature, speed, stiffness) twelve insertions were conducted. Measurements included six full insertions and six paused insertions. Paused insertions include a ten second paused time interval without further insertion movement each five millimetres. Measurements showed that dynamic and static components of the measured summed forces can be identified. Dynamic force components increase with increased insertion speeds and also with increased stiffness of the EC substitutes. Both force components decrease with larger radius of the PTFE model. After the insertion, the EC substitutes showed a curved shape, which indicates a plastic deformation of the embedded wires due to the insertion into the curved models. The results can be used for further research on an analytical model to predict the insertions forces of a specific combination of selected parameters as insertion speed and type of EC, combined with given factors such as cochlear geometry.

scholarly journals Nanoscopic changes in the lattice structure of striated muscle sarcomeres involved in the mechanism of spontaneous oscillatory contraction (SPOC)

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Fumiaki Kono ◽  
Seitaro Kawai ◽  
Yuta Shimamoto ◽  
Shin’ichi Ishiwata
Keyword(s):  
Structural Changes ◽  
Striated Muscle ◽  
Lattice Structure ◽  
Three Dimensional ◽  
Force Balance ◽  
Dynamic Force ◽  
Structural Requirement ◽  
Oscillatory Dynamics ◽  
Contractile System ◽  
Wide Range

Abstract Muscles perform a wide range of motile functions in animals. Among various types are skeletal and cardiac muscles, which exhibit a steady auto-oscillation of force and length when they are activated at an intermediate level of contraction. This phenomenon, termed spontaneous oscillatory contraction or SPOC, occurs devoid of cell membranes and at fixed concentrations of chemical substances, and is thus the property of the contractile system per se. We have previously developed a theoretical model of SPOC and proposed that the oscillation emerges from a dynamic force balance along both the longitudinal and lateral axes of sarcomeres, the contractile units of the striated muscle. Here, we experimentally tested this hypothesis by developing an imaging-based analysis that facilitates detection of the structural changes of single sarcomeres at unprecedented spatial resolution. We found that the sarcomere width oscillates anti-phase with the sarcomere length in SPOC. We also found that the oscillatory dynamics can be altered by osmotic compression of the myofilament lattice structure of sarcomeres, but they are unchanged by a proteolytic digestion of titin/connectin—the spring-like protein that provides passive elasticity to sarcomeres. Our data thus reveal the three-dimensional mechanical dynamics of oscillating sarcomeres and suggest a structural requirement of steady auto-oscillation.

scholarly journals Hepatitis C virus promotes virion secretion through cleavage of the Rab7 adaptor protein RILP

2016 ◽  
Vol 113 (44) ◽  
pp. 12484-12489 ◽  
Author(s):  
Ann L. Wozniak ◽  
Abby Long ◽  
Kellyann N. Jones-Jamtgaard ◽  
Steven A. Weinman
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Time Course ◽  
Rna Virus ◽  
Adaptor Protein ◽  
Virus Production ◽  
Hcv Rna ◽  
Cell Periphery ◽  
Dynein Motor ◽  
Intracellular Virus

Hepatitis C virus (HCV) is an enveloped RNA virus that modifies intracellular trafficking processes. The mechanisms that HCV and other viruses use to modify these events are poorly understood. In this study, we observed that two different RNA viruses, HCV and Sendai, cause inhibition of ras-related protein Rab-7 (Rab7)-dependent endosome–lysosome fusion. In both cases, viral infection causes cleavage of the Rab7 adaptor protein RILP (Rab interacting lysosomal protein), which is responsible for linking Rab7 vesicles to dynein motor complexes. RILP cleavage results in the generation of a cleaved RILP fragment (cRILP) missing the N terminus of the molecule. Although RILP localizes in a perinuclear fashion, cRILP moves to the cell periphery. Both knockdown of RILP and expression of cRILP reproduced the HCV-induced trafficking defect, and restoring full-length RILP reversed the trafficking effects of virus. For the first 3 d after electroporation of HCV RNA, intracellular virus predominates over secreted virus, but the quantity of intracellular virus then rapidly declines as secreted virus dominates. The transition from the intracellular-predominant to the secretion-predominant phenotype corresponds to the time course of cRILP generation. Expressing cRILP directly prevents intracellular virus accumulation at early times without affecting net virus production. The ability of cRILP to promote virus secretion could be prevented by a kinesin inhibitor. HCV thus modifies cellular trafficking by cleaving RILP, which serves to redirect Rab7-containing vesicles to a kinesin-dependent trafficking mode promoting virion secretion. Cleavage of a Rab adaptor protein is thus a mechanism by which viruses modify trafficking patterns of infected cells.

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