Actin‐based phagosome motility

Zhang, Fangliang; Southwick, Frederick S.; Purich, Daniel L.

doi:10.1002/cm.10058

Cited by 37 publications

(30 citation statements)

References 27 publications

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“…Occasionally, we saw actin comet/tail formation on endocytic organelles, in agreement with Merrifield et al (1999), Rozelle et al (2000), and Taunton et al (2000) (Figure 3E). Although comets has also been reported on LBP phagosomes in vivo (Zhang et al, 2002;Southwick et al, 2003), we never observed these on LBP phagosomes in our in vitro system. Thus, from the time the incubation is initiated until the 30-min time point when actin polymerization ceases and the viscoelasticity increases exponentially (Jahraus et al, 2001), there was a demonstrable emergence of an organization in the actin cytoskeleton, whose assembly is under the direct control of the cellular membrane organelles.…”

contrasting

confidence: 69%

Fusion between Phagosomes, Early and Late Endosomes: A Role for Actin in Fusion between Late, but Not Early Endocytic Organelles

Kjeken

Egeberg

Habermann

et al. 2004

MBoC

107

101

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Actin is implicated in membrane fusion, but the precise mechanisms remain unclear. We showed earlier that membrane organelles catalyze the de novo assembly of F-actin that then facilitates the fusion between latex bead phagosomes and a mixture of early and late endocytic organelles. Here, we correlated the polymerization and organization of F-actin with phagosome and endocytic organelle fusion processes in vitro by using biochemistry and light and electron microscopy. When membrane organelles and cytosol were incubated at 37°C with ATP, cytosolic actin polymerized rapidly and became organized into bundles and networks adjacent to membrane organelles. By 30-min incubation, a gel-like state was formed with little further polymerization of actin thereafter. Also during this time, the bulk of in vitro fusion events occurred between phagosomes/endocytic organelles. The fusion between latex bead phagosomes and late endocytic organelles, or between late endocytic organelles themselves was facilitated by actin, but we failed to detect any effect of perturbing F-actin polymerization on early endosome fusion. Consistent with this, late endosomes, like phagosomes, could nucleate F-actin, whereas early endosomes could not. We propose that actin assembled by phagosomes or late endocytic organelles can provide tracks for fusion-partner organelles to move vectorially toward them, via membranebound myosins, to facilitate fusion. INTRODUCTIONActin is essential for many cellular processes and dynamic polymerization/depolymerization of actin filaments is a critical property of all eukaryotic cells (Mitchison and Cramer, 1996;Carlier, 1998;Machesky and Insall, 1999;Small et al., 1999;Amann and Pollard, 2000). Much of the F-actin assembled in cells is intimately associated with membranes, especially the plasma membrane (Tilney, 1976;Hoglund et al., 1980;Lindberg et al., 1981;Carraway and Carraway, 1989;Dickinson and Purich, 2002). The first links between F-actin and membranes were reported by Tilney and Cardell (1970) who demonstrated the role of plasma membranes in actin nucleation, and Orci et al. (1972) who showed that cytochalasin can either stimulate or inhibit exocytic fusion, depending on the conditions. In addition to exocytosis (Bernstein et al., 1998;Lang et al., 2000;Bader et al., 2002), actin is now known to be involved in many other trafficking events, including phagocytosis (Swanson et al., 1999; Desjardin and Griffiths, 2003) and transcytosis (Durrbach et al., 2000). In the endocytic pathway, actin and myosins are essential for two different transport steps: clathrin-dependent internalization from the plasma membrane and transport to lysosomes (Durrbach et al., 1996;Riezman et al., 1996;Buss et al., 2001). More recently, actin has also been shown to be directly involved in homotypic fusion between yeast vacuoles (see DISCUSSION).Our interest in actin emerged from our analysis of an in vitro phagosome-endocytic organelle fusion assay that used latex bead phagosomes (LBP) and early endosomes (EE) and late endocytic org...

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contrasting

confidence: 69%

Fusion between Phagosomes, Early and Late Endosomes: A Role for Actin in Fusion between Late, but Not Early Endocytic Organelles

Kjeken

Egeberg

Habermann

et al. 2004

MBoC

107

101

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“…Furthermore, Arp2/3 complex is present throughout the actin tail of endosomes and vesicles (Rozelle et al, 2000;Taunton et al, 2000;Southwick et al, 2003) and colocalizes with flashing actin. Actin-based phagosome motility in macrophages does not depend on the surface coating on the bead (Zhang et al, 2002). Likewise, flashing does not depend on the molecular features of the internalized object.…”

Section: Repeated Cycles Of Actin Assembly and Disassembly Occurs On mentioning

confidence: 90%

“…All are transient events and, under unstimulated conditions, infrequent events. Actin tails on macrophage phagosomes persist for only 1-2 min (Zhang et al, 2002). Actin tails on vesicles induced by PI-5 kinase overexpression persist for an average of 2.2 min (Rozelle et al, 2000).…”

Section: Repeated Cycles Of Actin Assembly and Disassembly Occurs On mentioning

confidence: 98%

“…Actin comet tails have been observed in vivo on endosomes and lysosomes stimulated with phorbol 12-myristate 13-acetate (Taunton et al, 2000), endosomes in macrophages treated with lanthanum and zinc ions (Southwick et al, 2003), pinosomes (Merrifield et al, 1999), and Golgi and endocytic vesicles in cells overexpressing phosphatidylinositol phosphate 5-kinase (PI 5-kinase) or treated with pervanadate and platelet-derived growth factor (Rozelle et al, 2000). In addition, actin-based phagosome motility has been observed at a very low frequency (0.6 -1.5% of engulfed beads over a 10-min interval) in unstimulated macrophages (Zhang et al, 2002).…”

Section: Repeated Cycles Of Actin Assembly and Disassembly Occurs On mentioning

confidence: 99%

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Repeated Cycles of Rapid Actin Assembly and Disassembly on Epithelial Cell Phagosomes

Yam

Theriot

2004

MBoC

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We have found that early in infection of the intracellular pathogen Listeria monocytogenes in Madin-Darby canine kidney epithelial cells expressing actin conjugated to green fluorescent protein, F-actin rapidly assembles (ϳ25 s) and disassembles (ϳ30 s) around the bacteria, a phenomenon we call flashing. L. monocytogenes strains unable to perform actin-based motility or unable to escape the phagosome were capable of flashing, suggesting that the actin assembly occurs on the phagosome membrane. Cycles of actin assembly and disassembly could occur repeatedly on the same phagosome. Indirect immunofluorescence showed that most bacteria were fully internalized when flashing occurred, suggesting that actin flashing does not represent phagocytosis. Escherichia coli expressing invA, a gene product from Yersinia pseudotuberculosis that mediates cellular invasion, also induced flashing. Furthermore, polystyrene beads coated with E-cadherin or transferrin also induced flashing after internalization. This suggests that flashing occurs downstream of several distinct molecular entry mechanisms and may be a general consequence of internalization of large objects by epithelial cells. INTRODUCTIONBacterial pathogens such as Salmonella, Yersinia, and Listeria have a variety of mechanisms that allow them to enter and survive in eukaryotic cells. Their ability to survive intracellularly can protect them from host defenses and allow them to cross epithelial barriers. To enter eukaryotic cells, these invasive bacteria actively induce their own uptake by phagocytosis in normally nonphagocytic cells. Internalized bacteria either survive within the phagosome, encapsulated by the host cell membrane, or they escape from the phagosome and disseminate throughout the cytoplasm and to neighboring cells (reviewed in Cossart and Sansonetti, 2004). Modulation of the host cell actin cytoskeleton by invasive bacteria allows them to control many of these steps, including attachment and entry into cells, survival in the phagosome, and movement in the cytoplasm.One bacterium with an intracellular niche is Listeria monocytogenes, a food-borne pathogen responsible for meningitis, meningoencephalitis, septicemia, gastroenteritis, and abortions (reviewed in Cossart and Lecuit, 1998). Two L. monocytogenes proteins, internalin (InlA) and internalin B (InlB), can interact with the host cell surface proteins E-cadherin, gC1q-R, and Met to induce bacterial uptake into nonphagocytic cells (Mengaud et al., 1996;Braun et al., 2000;Shen et al., 2000). Uptake of bacteria is driven by actin polymerization and membrane extension (reviewed in Cossart and Sansonetti, 2004). Once inside the cell, L. monocytogenes escapes from the phagosome into the cytoplasm. In the cytoplasm, L. monocytogenes nucleates actin filaments that assemble into actin clouds around the bacteria and then into polarized comet tails. The actin comet tails propel the movement of L. monocytogenes within the cytoplasm of an infected cell and into neighboring cells (Tilney and Portnoy, 1989;Mounier e...

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“…This process occasionally initiated directed movement, and the link between flashing and actin comet rocketing was suggested. Nevertheless, with the exception of actin comets assembling on intracytoplasmic pathogens that have been extensively described (Gouin et al, 1999;Cudmore et al, 1995), it has been difficult to assign any function to actin-driven motility of the host cell endosomes (Taunton et al, 2000), pinosomes (Merrifield et al, 1999) or phagosomes (Zhang et al, 2002;Clarke et al, 2006).…”

Section: Introductionmentioning

confidence: 99%

Transient assembly of F-actin by phagosomes delays phagosome fusion with lysosomes in cargo-overloaded macrophages

Liebl¹,

Griffiths²

2009

Journal of Cell Science

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Dynamic remodelling of the cortical actin cytoskeleton is required for phagocytic uptake of pathogens and other particles by macrophages. Actin can also be nucleated de novo on membranes of nascent phagosomes, a process that can stimulate or inhibit phagosome fusion with lysosomes. Recently, phagosomes were shown to polymerize actin in transient pulses, called actin 'flashing', whose function remains unexplained. Here, we investigated phagosomal actin dynamics in live macrophages expressing actin tagged with green fluorescent protein (GFP). We show that only immature phagosomes can transiently induce assembly of actin coat, which forms a barrier preventing phagosome-lysosome docking and fusion. The capacity of phagosomes to assemble actin is enhanced in cells exposed to increased phagocytic load, which also exhibit a delay in phagosome maturation. Parallel analysis indicated that polymerization of actin on macropinosomes also induces compression and propulsion. We show that dynamic interactions between membrane elastic tension and compression forces of polymerizing actin can also lead to macropinosome constriction and scission -a process that is obstructed on rigid phagosomes. We hypothesize that the rate of individual phagosome maturation, as well as the biogenesis and remodelling of macropinosomes, can be regulated by the extent and manner of actin assembly on their membrane. Supplementary material available online at

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Actin‐based phagosome motility

Cited by 37 publications

References 27 publications

Fusion between Phagosomes, Early and Late Endosomes: A Role for Actin in Fusion between Late, but Not Early Endocytic Organelles

Fusion between Phagosomes, Early and Late Endosomes: A Role for Actin in Fusion between Late, but Not Early Endocytic Organelles

Repeated Cycles of Rapid Actin Assembly and Disassembly on Epithelial Cell Phagosomes

Transient assembly of F-actin by phagosomes delays phagosome fusion with lysosomes in cargo-overloaded macrophages

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