Interaction of Human Arp2/3 Complex and the
            <i>Listeria monocytogenes</i>
            ActA Protein in Actin Filament Nucleation

Welch, Matthew D.; Rosenblatt, Jody; Skoble, Justin; Portnoy, Daniel A.; Mitchison, Timothy J.

doi:10.1126/science.281.5373.105

Cited by 481 publications

(442 citation statements)

References 38 publications

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“…Remarkably, subsequent work (16)(17)(18) showed that Arp2/ 3 complex produces filaments with the properties of this model (Fig. 4).…”

Section: Mogilner and Oster 1996: Elastic Brownian Ratchet Mechanismsupporting

confidence: 58%

Theory from the Oster Laboratory Leaps Ahead of Experiment in Understanding Actin-Based Cellular Motility

Pollard

2016

Biophysical Journal

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By the early 1990s, most biophysicists and cell biologists agreed that polymerization of actin filaments at the leading edge of a motile cell pushes the plasma membrane forward, creating a protrusion that extends the border of the cell (Fig. 1). For cells on a flat substrate, like a microscope slide, this leading edge is a relatively flat lamella less than 1 mm thick. To balance protrusion at the front end, contractility of the cytoplasm was believed to pull the rear of the cell forward toward attachments that form continuously behind the leading edge.Multiple lines of evidence supported the idea that actin polymerization drives cellular movements. Early electron micrographs of thin sections showed microfilaments (subsequently confirmed to be actin) in the cytoplasm next to the leading edge (1). Even better, electron micrographs of negatively stained specimens showed dense arrays of filaments at angles to the plasma membrane (2). Drugs that interfere with actin polymerization stop the protrusion of the leading edge (3), and two elegant fluorescence microscopy experiments established that actin polymerizes near the leading edge. First, Yu-Li Wang injected live cells with fluorescent actin, which incorporated into cellular actin structures (4). When he bleached a spot in the fluorescent leading lamella of the stationary cell, the spot moved away from the leading edge as new fluorescent actin was incorporated next to the plasma membrane. Over time, the fluorescence recovered, showing that the filaments turned over. Julie Theriot and Tim Mitchison used photoactivation to learn more (5). They injected motile cells with actin coupled to a ''caged'' fluorescent dye. After the tagged actin molecules incorporated into cellular actin structures, they uncaged the fluorescent dye with a light pulse near the leading edge. As the cell moved forward, the spot of fluorescent actin was stationary relative to the substrate and turned over on a time scale of tens of seconds. Electron microscopy (2) showed that these leading edge actin filaments are mostly oriented with their faster growing ''barbed ends'' (6) toward the leading edge of motile cells.Many of these ideas were reinforced by parallel experiments on the movements of certain bacteria through the cytoplasm of host animal cells (7). Actin polymerization next to the bacterium assembles a comet tail of actin filaments that propels the bacterium through the cytoplasm (8,9). This work culminated in reconstitution of bacterial motility from purified actin and a few other proteins (10).Knowing the equilibrium constant for subunit addition to actin filaments, Hill and Kirschner made solid thermodynamic arguments for how polymerization might produce the force to push the membrane forward (11). However, Peskin, Odell, and Oster (12) pointed out ''such arguments provide no mechanistic explanation of how the free energy of polymerization is actually transduced into directed mechanical force.'' A series of three classic papers in Biophysical Journal by George Oster with Charles Peskin...

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“…Remarkably, subsequent work (16)(17)(18) showed that Arp2/ 3 complex produces filaments with the properties of this model (Fig. 4).…”

Section: Mogilner and Oster 1996: Elastic Brownian Ratchet Mechanismsupporting

confidence: 58%

Theory from the Oster Laboratory Leaps Ahead of Experiment in Understanding Actin-Based Cellular Motility

Pollard

2016

Biophysical Journal

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“…Actin cytoskeleton has been shown to play a role in the infection of all the viruses tested in this study according to previous studies using the chemical actin inhibitor (Rosenthal et al, 1985;Kizhatil and Albritton, 1997;Bukrinskaya et al, 1998;Iyengar et al, 1998;Li et al, 1998;Locker et al, 2000). In addition, the Arp2/3 complex-mediated actin nucleation is sensitive to CC (Welch et al, 1998). Therefore, GFP-VCA's ability to limit infection of primate lentiviruses and IMV might be a part of, if not all, the mechanism by which CC reduced the efficiency of viral infection.…”

Section: Discussionmentioning

confidence: 91%

Inhibiting the Arp2/3 Complex Limits Infection of Both Intracellular Mature Vaccinia Virus and Primate Lentiviruses

Komano

Miyauchi

Matsuda

et al. 2004

MBoC

100

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Characterizing cellular factors involved in the life cycle of human immunodeficiency virus type 1 (HIV-1) is an initial step toward controlling replication of HIV-1. Actin polymerization mediated by the Arp2/3 complex has been found to play a critical role in some pathogens' intracellular motility. We have asked whether this complex also contributes to the viral life cycles including that of HIV-1. We have used both the acidic domains from actin-related protein (Arp) 2/3 complex-binding proteins such as the Wiscott-Aldrich syndrome protein (N-WASP) or cortactin, and siRNA directing toward Arp2 to inhibit viral infection. HIV-1, simian immunodeficiency virus (SIV), and intracellular mature vaccinia virus (IMV) were sensitive to inhibition of the Arp2/3 complex, whereas MLV, HSV-1, and adenovirus were not. Interestingly, pseudotyping HIV-1 with vesicular stomatitis virus G protein (VSV-G) overcame this inhibition. Constitutive inhibition of the Arp2/3 complex in the T-cell line H9 also blocked replication of HIV-1. These data suggested the existence of an Arp2/3 complex-dependent event during the early phase of the life cycles of both primate lentiviruses and IMV. Inhibiting the HIV-1's ability to activate Arp2/3 complex could be a potential chemotherapeutic intervention for acquired immunodeficiency syndrome (AIDS)

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“…In contrast, WASP and WAVE1 did not accumulate near the bacterial surface. We also examined the ability of Listeria monocytogenes to recruit WASP family proteins, as Listeria induces actin assembly in infected cells that is mediated by ActA through binding and stimulating the Arp2/3 complex (Welch et al, 1998). Consequently, none of the WASP family proteins including N-WASP associated with the surface of Listeria (data not shown), thus suggesting that Listeria motility does not depend on the presence of WASP family proteins.…”

Section: Shigella Virg Binds To N-wasp But Not To the Other Wasp Famimentioning

confidence: 99%

Neural Wiskott-Aldrich syndrome protein (N-WASP) is the specific ligand for Shigella VirG among the WASP family and determines the host cell type allowing actin-based spreading

et al. 2002

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SummaryShigella, the causative agent of bacillary dysentery, is capable of directing its movement within host cells by forming an actin comet tail. The VirG (IcsA) protein expressed at one pole of the bacterium recruits neural Wiskott-Aldrich syndrome protein (N-WASP), a member of the WASP family, which in turn stimulates actin-related protein (Arp) 2/3 complex-mediated actin polymerization. As all the WASP family proteins induce actin polymerization by recruiting Arp2/3 complex, we investigated their involvement in Shigella motility. Here, we show that VirG binds to N-WASP but not to the other WASP family proteins. Using a series of chimeras obtained by swapping N-WASP and WASP domains, we demonstrated that the specificity of VirG to interact with N-WASP lies in the N-terminal region containing the pleckstrin homology (PH) domain and calmodulin-binding IQ motif of N-WASP. A conformational change in N-WASP was important for the VirG-N-WASP interaction, as elimination of the C-terminal acidic region, which is responsible for the intramolecular interaction with the central basic region of N-WASP, affected the specific binding to VirG. We observed that, in haematopoietic cells such as macrophages, polymorphonuclear leucocytes (PMNs) and platelets, WASP was predominantly expressed, whereas the expression of N-WASP was greatly suppressed. Indeed, unlike Listeria, Shigella was unable to move in macrophages at all, (V) domain, a domain (C) with homology to the actindepolymerizing protein cofilin and, finally, a C-terminal acidic (A) segment (Miki et al., 1996). The C-terminal VCA domain interacts with the actin-related protein (Arp) 2/3 complex after the stimulation of actin polymerization by the complex (Rohatgi et al., 1999). N-WASP exists in two conformations: an inactive form in which the VCA domain is masked, and an active form in which it is exposed to promote actin polymerization by interaction with the Arp2/3 complex. The activation of N-WASP requires Cdc42 or PtdIns(4,5)P2 bound to N-WASP (Miki et al., 1998b;Rohatgi et al., 1999;2000;Prehoda et al., 2000). It has been reported that N-WASP is required for generating the actin tail from motile S. flexneri (Suzuki et al., 1998;Egile et al., 1999). Arp2/3 complex is also required for actin-based motility of Shigella, in which N-WASP stimulates the actin nucleation activity of the Arp2/3 complex in vitro (Egile et al., 1999).In mammalian cells, five WASP family members, N-WASP, WASP and WAVE1-3, have been identified and characterized to date (Suetsugu et al., 1999); however, none of them except N-WASP has been elucidated for involvement in the actin-based motility of Shigella in infected cells. It has been shown that human platelet extracts supported Shigella actin-based motility as added N-WASP in vitro (Egile et al., 1999). Recent reports using N-WASP-deficient mouse embryonic fibroblasts (Lommel et al., 2001;Snapper et al., 2001) confirmed our previous work indicating that N-WASP is required for Shigella motility in mammalian cells (Suzuki et al., 1998). However...

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Interaction of Human Arp2/3 Complex and the Listeria monocytogenes ActA Protein in Actin Filament Nucleation

Cited by 481 publications

References 38 publications

Theory from the Oster Laboratory Leaps Ahead of Experiment in Understanding Actin-Based Cellular Motility

Theory from the Oster Laboratory Leaps Ahead of Experiment in Understanding Actin-Based Cellular Motility

Inhibiting the Arp2/3 Complex Limits Infection of Both Intracellular Mature Vaccinia Virus and Primate Lentiviruses

Neural Wiskott-Aldrich syndrome protein (N-WASP) is the specific ligand for Shigella VirG among the WASP family and determines the host cell type allowing actin-based spreading

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