Choosing orientation: Influence of cargo geometry and ActA polarization on actin comet tails

Department of Biochemistry, Stanford University School of Medicine, Stanford, CA 94305, USA.
Molecular biology of the cell (Impact Factor: 4.47). 01/2012; 23(4):614-29. DOI: 10.1091/mbc.E11-06-0584
Source: PubMed


Networks of polymerizing actin filaments can propel intracellular pathogens and drive movement of artificial particles in reconstituted systems. While biochemical mechanisms activating actin network assembly have been well characterized, it remains unclear how particle geometry and large-scale force balance affect emergent properties of movement. We reconstituted actin-based motility using ellipsoidal beads resembling the geometry of Listeria monocytogenes. Beads coated uniformly with the L. monocytogenes ActA protein migrated equally well in either of two distinct orientations, with their long axes parallel or perpendicular to the direction of motion, while intermediate orientations were unstable. When beads were coated with a fluid lipid bilayer rendering ActA laterally mobile, beads predominantly migrated with their long axes parallel to the direction of motion, mimicking the orientation of motile L. monocytogenes. Generating an accurate biophysical model to account for our observations required the combination of elastic-propulsion and tethered-ratchet actin-polymerization theories. Our results indicate that the characteristic orientation of L. monocytogenes must be due to polarized ActA rather than intrinsic actin network forces. Furthermore, viscoelastic stresses, forces, and torques produced by individual actin filaments and lateral movement of molecular complexes must all be incorporated to correctly predict large-scale behavior in the actin-based movement of nonspherical particles.

Download full-text


Available from: Frederick Soo, Feb 04, 2015
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The cytoskeleton architecture supports many cellular functions. Cytoskeleton networks form complex intracellular structures that vary during the cell cycle and between different cell types according to their physiological role. These structures do not emerge spontaneously. They result from the interplay between intrinsic self-organization properties and the conditions imposed by spatial boundaries. Along these boundaries, cytoskeleton filaments are anchored, repulsed, aligned, or reoriented. Such local effects can propagate alterations throughout the network and guide cytoskeleton assembly over relatively large distances. The experimental manipulation of spatial boundaries using microfabrication methods has revealed the underlying physical processes directing cytoskeleton self-organization. Here we review, step-by-step, from molecules to tissues, how the rules that govern assembly have been identified. We describe how complementary approaches, all based on controlling geometric conditions, from in vitro reconstruction to in vivo observation, shed new light on these fundamental organizing principles.
    Trends in cell biology 09/2012; 22(12). DOI:10.1016/j.tcb.2012.08.012 · 12.01 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Two theoretical models dominate current understanding of actin-based propulsion: microscopic polymerization ratchet model predicts that growing and writhing actin filaments generate forces and movements, while macroscopic elastic propulsion model suggests that deformation and stress of growing actin gel are responsible for the propulsion. We examine both experimentally and computationally the 2D movement of ellipsoidal beads propelled by actin tails and show that neither of the two models can explain the observed bistability of the orientation of the beads. To explain the data, we develop a 2D hybrid mesoscopic model by reconciling these two models such that individual actin filaments undergoing nucleation, elongation, attachment, detachment and capping are embedded into the boundary of a node-spring viscoelastic network representing the macroscopic actin gel. Stochastic simulations of this 'in silico' actin network show that the combined effects of the macroscopic elastic deformation and microscopic ratchets can explain the observed bistable orientation of the actin-propelled ellipsoidal beads. To test the theory further, we analyze observed distribution of the curvatures of the trajectories and show that the hybrid model's predictions fit the data. Finally, we demonstrate that the model can explain both concave-up and concave-down force-velocity relations for growing actin networks depending on the characteristic time scale and network recoil. To summarize, we propose that both microscopic polymerization ratchets and macroscopic stresses of the deformable actin network are responsible for the force and movement generation.
    PLoS Computational Biology 11/2012; 8(11):e1002764. DOI:10.1371/journal.pcbi.1002764 · 4.62 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Many virulence factors of Gram-positive bacteria are anchored to the peptidoglycan by a sorting signal. While surface display mechanisms are well characterized, less is known about the spatial and temporal organization of these proteins in the bacterial envelope. This review summarizes recent studies on the rod-shaped Listeria monocytogenes, ovococcal Streptococcus pyogenes and spherical Staphylococcus aureus bacteria that provide insights into the compartmentalization of the surface and distribution of peptidoglycan-anchored proteins in space and time. We discuss models that support mechanistic bases for localization of proteins at the poles, septum or lateral sites. The results indicate that deployment of virulence factors by pathogenic bacteria is a dynamic process tightly connected to secretion, cell morphogenesis, cell division rate and gene expression levels.
    Current opinion in microbiology 11/2012; 15(6). DOI:10.1016/j.mib.2012.10.010 · 5.90 Impact Factor
Show more