LIM proteins in actin cytoskeleton mechanoresponse

pre-printThe actin cytoskeleton assembles into branched networks or bundles to generate mechanical force for critical cellular processes such as establishment of polarity, adhesion, and migration. Stress fibers are contractile, actomyosin structures that physically couple to the extracellular matrix through integrin-based focal adhesions, thereby transmitting force into and across the cell. Recently, LIM domain proteins have been implicated in mediating this cytoskeletal mechanotransduction. Among the more well studied LIM domain adapter proteins is zyxin, a dynamic component of both focal adhesions and stress fibers. Here, we discuss recent research detailing the mechanisms by which stress fibers adjust their structure and composition to balance mechanical forces, and suggest ways zyxin and other LIM domain proteins mediate mechanoresponse

Mathematical modeling of the dynamic mechanical behavior of neighboring sarcomeres in actin stress fibers

pre-printActin stress fibers (SFs) in live cells consist of series of dynamic individual sarcomeric units. Within a group of consecutive SF sarcomeres, individual sarcomeres can spontaneously shorten or lengthen without changing the overall length of this group, but the underlying mechanism is unclear. We used a computational model to test our hypothesis that this dynamic behavior is inherent to the heterogeneous mechanical properties of the sarcomeres and the cytoplasmic viscosity. Each sarcomere was modeled as a discrete element consisting of an elastic spring, a viscous dashpot and an active contractile unit all connected in parallel, and experiences forces as a result of actin filament elastic stiffness, myosin II contractility, internal viscoelasticity, or cytoplasmic drag. When all four types of forces are considered, the simulated dynamic behavior closely resembles the experimental observations, which include a low-frequency fluctuation in individual sarcomere length and compensatory lengthening and shortening of adjacent sarcomeres. Our results suggest that heterogeneous stiffness and viscoelasticity of actin fibers, heterogeneous myosin II contractility, and the cytoplasmic drag are sufficient to cause spontaneous fluctuations in SF sarcomere length. Our results shed new light to the dynamic behavior of SF and help design experiments to further our understanding of SF dynamics

Mutations in Drosophila enabled and rescue by human vasodilator-stimulated phosphoprotein (VASP) indicate important functional roles for Ena/VASP homology domain 1 (EVH1) and EVH2 domains

Journal ArticleDrosophila Enabled (Ena) was initially identified as a dominant genetic suppressor of mutations in the Abelson tyrosine kinase and, more recently, as a member of the Ena/human vasodilator-stimulated phosphoprotein (VASP) family of proteins. We have used genetic, biochemical, and cell biological approaches to demonstrate the functional relationship between Ena and human VASP

Effective free energy for pinned membranes

We consider membranes adhered through specific receptor-ligand bonds. Thermal undulations of the membrane induce effective interactions between adhesion sites. We derive an upper bound to the free energy that is independent of interaction details. To lowest order in a systematic expansion we obtain two-body interactions which allow to map the free energy onto a lattice gas with constant density. The induced interactions alone are not strong enough to lead to a condensation of individual adhesion sites. A measure of the thermal roughness is shown to depend on the inverse square root of the density of adhesion sites, which is in good agreement with previous computer simulations.Comment: to appear as a Rapid Communication in Phys. Rev.

Molecular dissection of the mechanism by which EWS/FLI expression compromises actin cytoskeletal integrity and cell adhesion in Ewing sarcoma.

Ewing sarcoma is the second-most-common bone cancer in children. Driven by an oncogenic chromosomal translocation that results in the expression of an aberrant transcription factor, EWS/FLI, the disease is typically aggressive and micrometastatic upon presentation. Silencing of EWS/FLI in patient-derived tumor cells results in the altered expression of hundreds to thousands of genes and is accompanied by dramatic morphological changes in cytoarchitecture and adhesion. Genes encoding focal adhesion, extracellular matrix, and actin regulatory proteins are dominant targets of EWS/FLI-mediated transcriptional repression. Reexpression of genes encoding just two of these proteins, zyxin and α5 integrin, is sufficient to restore cell adhesion and actin cytoskeletal integrity comparable to what is observed when the EWS/FLI oncogene expression is compromised. Using an orthotopic xenograft model, we show that EWS/FLI-induced repression of α5 integrin and zyxin expression promotes tumor progression by supporting anchorage-independent cell growth. This selective advantage is paired with a tradeoff in which metastatic lung colonization is compromised

A Zyxin-Mediated Mechanism for Actin Stress Fiber Maintenance and Repair

SummaryTo maintain mechanical homeostasis, cells must recognize and respond to changes in cytoskeletal integrity. By imaging live cells expressing fluorescently tagged cytoskeletal proteins, we observed that actin stress fibers undergo local, acute, force-induced elongation and thinning events that compromise their stress transmission function, followed by stress fiber repair that restores this capability. The LIM protein zyxin rapidly accumulates at sites of strain-induced stress fiber damage and is essential for stress fiber repair and generation of traction force. Zyxin promotes recruitment of the actin regulatory proteins α-actinin and VASP to compromised stress fiber zones. α-Actinin plays a critical role in restoration of actin integrity at sites of local stress fiber damage, whereas both α-actinin and VASP independently contribute to limiting stress fiber elongation at strain sites, thus promoting stabilization of the stress fiber. Our findings demonstrate a mechanism for rapid repair and maintenance of the structural integrity of the actin cytoskeleton

Evidence for Different Freeze-Out Radii of High- and Low-Energy Pions Emitted in Au+Au Collisions at 1 GeV/nucleon

Double differential production cross sections of negative and positive pions and the number of participating protons have been measured in central Au+Au collisions at 1 GeV per nucleon incident energy. At low pion energies the pi^- yield is strongly enhanced over the pi^+ yield. The energy dependence of the pi^-/pi^+ ratio is assigned to the Coulomb interaction of the charged pions with the protons in the reaction zone. The deduced Coulomb potential increases with increasing pion c.m. energy. This behavior indicates different freeze-out radii for different pion energies in the c.m.~frame.Comment: IKDA is the Institute for Nuclear Physics in Darmstadt/German