143 research outputs found
Fluorescence Correlation Spectroscopy analysis of segmental dynamics in Actin filaments
We adapt Fluorescence Correlation spectroscopy (FCS) formalism to the studies
of the dynamics of semi-flexible polymers and derive expressions relating FCS
correlation function to the longitudinal and transverse mean square
displacements of polymer segments. We use the derived expressions to measure
the dynamics of actin filaments in two experimental situations: filaments
labeled at distinct positions and homogeneously labeled filaments. Both
approaches give consistent results and allow to measure the temporal dependence
of the segmental mean-square displacement (MSD) over almost five decades in
time, from ~0.04ms to 2s. These noninvasive measurements allow for a detailed
quantitative comparison of the experimental data to the current theories of
semi-flexible polymer dynamics. Good quantitative agreement is found between
the experimental results and theories explicitly accounting for the
hydrodynamic interactions between polymer segments
Load fluctuations drive actin network growth
The growth of actin filament networks is a fundamental biological process
that drives a variety of cellular and intracellular motions. During motility,
eukaryotic cells and intracellular pathogens are propelled by actin networks
organized by nucleation-promoting factors, which trigger the formation of
nascent filaments off the side of existing filaments in the network. A Brownian
ratchet (BR) mechanism has been proposed to couple actin polymerization to
cellular movements, whereby thermal motions are rectified by the addition of
actin monomers at the end of growing filaments. Here, by following
actin--propelled microspheres using three--dimensional laser tracking, we find
that beads adhered to the growing network move via an object--fluctuating BR.
Velocity varies with the amplitude of thermal fluctuation and inversely with
viscosity as predicted for a BR. In addition, motion is saltatory with a broad
distribution of step sizes that is correlated in time. These data point to a
model in which thermal fluctuations of the microsphere or entire actin network,
and not individual filaments, govern motility. This conclusion is supported by
Monte Carlo simulations of an adhesion--based BR and suggests an important role
for membrane tension in the control of actin--based cellular protrusions.Comment: To be published in PNA
Thermodynamics and structure of self-assembled networks
We study a generic model of self-assembling chains which can branch and form
networks with branching points (junctions) of arbitrary functionality. The
physical realizations include physical gels, wormlike micells, dipolar fluids
and microemulsions. The model maps the partition function of a solution of
branched, self-assembling, mutually avoiding clusters onto that of a Heisenberg
magnet in the mathematical limit of zero spin components. The model is solved
in the mean field approximation. It is found that despite the absence of any
specific interaction between the chains, the entropy of the junctions induces
an effective attraction between the monomers, which in the case of three-fold
junctions leads to a first order reentrant phase separation between a dilute
phase consisting mainly of single chains, and a dense network, or two network
phases. Independent of the phase separation, we predict the percolation
(connectivity) transition at which an infinite network is formed that partially
overlaps with the first-order transition. The percolation transition is a
continuous, non thermodynamic transition that describes a change in the
topology of the system. Our treatment which predicts both the thermodynamic
phase equilibria as well as the spatial correlations in the system allows us to
treat both the phase separation and the percolation threshold within the same
framework. The density-density correlation correlation has a usual
Ornstein-Zernicke form at low monomer densities. At higher densities, a peak
emerges in the structure factor, signifying an onset of medium-range order in
the system. Implications of the results for different physical systems are
discussed.Comment: Submitted to Phys. Rev.
New Proposed Mechanism of Actin-Polymerization-Driven Motility
We present the first numerical simulation of actin-driven propulsion by
elastic filaments. Specifically, we use a Brownian dynamics formulation of the
dendritic nucleation model of actin-driven propulsion. We show that the model
leads to a self-assembled network that exerts forces on a disk and pushes it
with an average speed. This simulation approach is the first to observe a speed
that varies non-monotonically with the concentration of branching proteins
(Arp2/3), capping protein and depolymerization rate (ADF), in accord with
experimental observations. Our results suggest a new interpretation of the
origin of motility that can be tested readily by experiment.Comment: 31 pages, 5 figure
In Silico Reconstitution of Actin-Based Symmetry Breaking and Motility
Computational modeling and experimentation in a model system for actin-based force generation explain how actin networks initiate and maintain directional movement
Quantitative description of temperature induced self-aggregation thermograms determined by differential scanning calorimetry
A novel thermodynamic approach for the description of differential scanning calorimetry (DSC) experiments on self-aggregating systems is derived and presented. The method is based on a mass action model where temperature dependence of aggregation numbers is considered. The validity of the model was confirmed by describing the aggregation behavior of poly(ethylene oxide)-poly(propylene oxide) block copolymers, which are well-known to exhibit a strong temperature dependence. The quantitative description of the thermograms could be performed without any discrepancy between calorimetric and van 't Hoff enthalpies, and moreover, the aggregation numbers obtained from the best fit of the DSC experiments are in good agreement with those obtained by light scattering experiments corroborating the assumptions done in the derivation of the new model
An Experimental and Computational Study of the Effect of ActA Polarity on the Speed of Listeria monocytogenes Actin-based Motility
Listeria monocytogenes is a pathogenic bacterium that moves within infected cells and spreads directly between cells by harnessing the cell's dendritic actin machinery. This motility is dependent on expression of a single bacterial surface protein, ActA, a constitutively active Arp2,3 activator, and has been widely studied as a biochemical and biophysical model system for actin-based motility. Dendritic actin network dynamics are important for cell processes including eukaryotic cell motility, cytokinesis, and endocytosis. Here we experimentally altered the degree of ActA polarity on a population of bacteria and made use of an ActA-RFP fusion to determine the relationship between ActA distribution and speed of bacterial motion. We found a positive linear relationship for both ActA intensity and polarity with speed. We explored the underlying mechanisms of this dependence with two distinctly different quantitative models: a detailed agent-based model in which each actin filament and branched network is explicitly simulated, and a three-state continuum model that describes a simplified relationship between bacterial speed and barbed-end actin populations. In silico bacterial motility required a cooperative restraining mechanism to reconstitute our observed speed-polarity relationship, suggesting that kinetic friction between actin filaments and the bacterial surface, a restraining force previously neglected in motility models, is important in determining the effect of ActA polarity on bacterial motility. The continuum model was less restrictive, requiring only a filament number-dependent restraining mechanism to reproduce our experimental observations. However, seemingly rational assumptions in the continuum model, e.g. an average propulsive force per filament, were invalidated by further analysis with the agent-based model. We found that the average contribution to motility from side-interacting filaments was actually a function of the ActA distribution. This ActA-dependence would be difficult to intuit but emerges naturally from the nanoscale interactions in the agent-based representation
Interplay of Magnetic Interactions and Active Movements in the Formation of Magnetosome Chains
Magnetotactic bacteria assemble chains of magnetosomes, organelles that contain magnetic nano-crystals. A number of genetic factors involved in the controlled biomineralization of these crystals and the assembly of magnetosome chains have been identified in recent years, but how the specific biological regulation is coordinated with general physical processes such as diffusion and magnetic interactions remains unresolved. Here, these questions are addressed by simulations of different scenarios for magnetosome chain formation, in which various physical processes and interactions are either switched on or off. The simulation results indicate that purely physical processes of magnetosome diffusion, guided by their magnetic interactions, are not sufficient for the robust chain formation observed experimentally and suggest that biologically encoded active movements of magnetosomes may be required. Not surprisingly, the chain pattern is most resembling experimental results when both magnetic interactions and active movement are coordinated. We estimate that the force such active transport has to generate is compatible with forces generated by the polymerization or depolymerization of cytoskeletal filaments. The simulations suggest that the pleiotropic phenotypes of mamK deletion strains may be due to a defect in active motility of magnetosomes and that crystal formation in magneteosome vesicles is coupled to the activation of their active motility in M. gryphiswaldense, but not in M. magneticum
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