Arp2/3 complex interactions and actin network turnover in lamellipodia.

Cytoskeleton Dynamics Group, Helmholtz Centre for Infection Research, Braunschweig, Germany.
The EMBO Journal (Impact Factor: 9.82). 05/2008; 27(7):982-92. DOI: 10.1038/emboj.2008.34
Source: PubMed

ABSTRACT Cell migration is initiated by lamellipodia-membrane-enclosed sheets of cytoplasm containing densely packed actin filament networks. Although the molecular details of network turnover remain obscure, recent work points towards key roles in filament nucleation for Arp2/3 complex and its activator WAVE complex. Here, we combine fluorescence recovery after photobleaching (FRAP) of different lamellipodial components with a new method of data analysis to shed light on the dynamics of actin assembly/disassembly. We show that Arp2/3 complex is incorporated into the network exclusively at the lamellipodium tip, like actin, at sites coincident with WAVE complex accumulation. Capping protein likewise showed a turnover similar to actin and Arp2/3 complex, but was confined to the tip. In contrast, cortactin-another prominent Arp2/3 complex regulator-and ADF/cofilin-previously implicated in driving both filament nucleation and disassembly-were rapidly exchanged throughout the lamellipodium. These results suggest that Arp2/3- and WAVE complex-driven actin filament nucleation at the lamellipodium tip is uncoupled from the activities of both cortactin and cofilin. Network turnover is additionally regulated by the spatially segregated activities of capping protein at the tip and cofilin throughout the mesh.

  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Filopodia are sensors on both excitable and non-excitable cells. The sensing function is well documented in neurons and blood vessels of adult animals and is obvious during dorsal closure in embryonic development. Nerve cells extend neurites in a bidirectional fashion with growth cones at the tips where filopodia are concentrated. Their sensing of environmental cues underpins the axon's ability to "guide," bypassing non-target cells and moving toward the target to be innervated. This review focuses on the role of filopodia structure and dynamics in the detection of environmental cues, including both the extracellular matrix (ECM) and the surfaces of neighboring cells. Other protrusions including the stereocilia of the inner ear and epididymus, the invertebrate Type I mechanosensors, and the elongated processes connecting osteocytes, share certain principles of organization with the filopodia. Actin bundles, which may be inside or outside of the excitable cell, function to transduce stress from physical perturbations into ion signals. There are different ways of detecting such perturbations. Osteocyte processes contain an actin core and are physically anchored on an extracellular structure by integrins. Some Type I mechanosensors have bridge proteins that anchor microtubules to the membrane, but bundles of actin in accessory cells exert stress on this complex. Hair cells of the inner ear rely on attachments between the actin-based protrusions to activate ion channels, which then transduce signals to afferent neurons. In adherent filopodia, the focal contacts (FCs) integrated with ECM proteins through integrins may regulate integrin-coupled ion channels to achieve signal transduction. Issues that are not understood include the role of Ca(2+) influx in filopodia dynamics and how integrins coordinate or gate signals arising from perturbation of channels by environmental cues.
    Cellular Signalling 07/2013; · 4.47 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: While light microscopy allows for observations in live cells, structures spaced less than 200 nm are represented as a blur. Structured Illumination Microscopy (SIM) is a technique that is able to discern structures below this limit. Here we use SIM to visualize actin morphology at focal adhesions. Dual color super-resolution imaging reveals that mature adhesions display a dense packaging of parallel actin fibers, where active focal adhesion kinase associates with actin.
    BioSpektrum 18(4).
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: While the molecular and biophysical mechanisms underlying cell protrusion on two-dimensional substrates are well understood, our knowledge of the actin structures driving protrusion in three-dimensional environments is poor, despite relevance to inflammation, development and cancer. Here we report that, during chemotactic migration through microchannels with 5 μm × 5 μm cross-sections, HL60 neutrophil-like cells assemble an actin-rich slab filling the whole channel cross-section at their front. This leading edge comprises two distinct F-actin networks: an adherent network that polymerizes perpendicular to cell-wall interfaces and a 'free' network that grows from the free membrane at the cell front. Each network is polymerized by a distinct nucleator and, due to their geometrical arrangement, the networks interact mechanically. On the basis of our experimental data, we propose that, during interstitial migration, medial growth of the adherent network compresses the free network preventing its retrograde movement and enabling new polymerization to be converted into forward protrusion.
    Nature Communications 12/2013; 4:2896. · 10.74 Impact Factor

Full-text (2 Sources)

Available from
May 22, 2014