Listeria Protein ActA Mimics WASP Family Proteins:  It Activates Filament Barbed End Branching by Arp2/3 Complex

Boujemaa-Paterski, Rajaa; Gouin, Edith; Hansen, Gert H.; Samarin, Stanislav; Clainche, Christophe Le; Didry, Dominique; Dehoux, Pierre; Cossart, Pascale; Kocks, Christine; Mf, Carlier; Pantaloni, Dominique

doi:10.1021/bi010486b

Cited by 117 publications

(117 citation statements)

References 48 publications

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“…pY53-actin and unphosphorylated actin, at the indicated concentrations, containing 4% pyrene-labeled actin, with or without addition of spectrin-actin (36), gelsolin (Sigma)-actin seeds (37) or Arp2͞3-VCA (Cytoskeleton, Denver, CO) (38), was polymerized at room temperature in G-buffer by the addition of 100 mM KCl and 2 mM MgCl 2 (final concentrations). The increase in fluorescence was measured with excitation at 365 nm and emission at 407 nm.…”

Section: Methodsmentioning

confidence: 99%

Phosphorylation of actin Tyr-53 inhibits filament nucleation and elongation and destabilizes filaments

Liu¹,

Shu²,

Hong³

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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Dictyostelium actin was shown to become phosphorylated on Tyr-53 late in the developmental cycle and when cells in the amoeboid stage are subjected to stress but the phosphorylated actin had not been purified and characterized. We have separated phosphorylated and unphosphorylated actin and shown that Tyr-53 phosphorylation substantially reduces actin's ability to inactivate DNase I, increases actin's critical concentration, and greatly reduces its rate of polymerization. Tyr-53 phosphorylation substantially, if not completely, inhibits nucleation and elongation from the pointed end of actin filaments and reduces the rate of elongation from the barbed end. Negatively stained electron microscopic images of polymerized Tyr-53-phosphorylated actin show a variable mixture of small oligomers and filaments, which are converted to more typical, long filaments upon addition of myosin subfragment 1. Tyr-53-phosphorylated and unphosphorylated actin copolymerize in vitro, and phosphorylated and unphosphorylated actin colocalize in amoebae. Tyr-53 phosphorylation does not affect the ability of filamentous actin to activate myosin ATPase.actin polymerization ͉ Dictyostelium ͉ phosphorylated actin T ransfer of Dictyostelium amoebae from nutrient to nonnutrient medium initiates a 24-hour developmental cycle (1) in which the amoebae aggregate and differentiate to form multicellular organisms that mature to fruiting bodies containing stable spores from which, when they are placed in nutrient medium, amoebae germinate. Several laboratories (2-4) reported tyrosine phosphorylation of actin [phosphotyrosine actin (pY-actin)] correlated with rearrangements of the actin cytoskeleton during the developmental cycle. pY-actin appears late in maturing spores, i.e., Ϸ24 h into the developmental cycle, reaches a maximum level at Ϸ36 h, at which time Ϸ50% of the actin is phosphorylated (3), remains constant for Ϸ20 days, at 22°C, and then decreases, disappearing entirely by 30 days, at which time the spores are no longer viable (3). When viable spores are placed in nutrient medium, pY-actin is dephosphorylated, with a half-life of Ϸ5 min (3), before spore swelling and germination (2, 4).Although vegetative amoebae in nutrient medium contain little or no pY-actin (5, 6), phosphorylation transiently increases (for Ϸ20-25 min) when amoebae are transferred from nonnutrient to nutrient medium (7), with concurrent changes in cell shape, for example, loss of pseudopods, rounding up of the previously elongated cells, and weakened adherence to the substratum (7). Tyrosine phosphorylation also occurs when vegetative amoebae in nutrient medium are exposed to phenylarsine oxide (PAO) (5), an inhibitor of phosphotyrosine phosphatase, or are subjected to stress, for example, inhibition of oxidative phosphorylation (8, 9) or elevated temperature (9), with parallel changes in cell shape similar to the changes that occur when cells are transferred from nonnutrient to nutrient medium.Importantly, phosphorylation occurs uniquely at Tyr-53 (9). Thus, phosphoryl...

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Section: Methodsmentioning

confidence: 99%

Phosphorylation of actin Tyr-53 inhibits filament nucleation and elongation and destabilizes filaments

Liu¹,

Shu²,

Hong³

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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“…163 The N-terminal region of ActA is sufficient for its activity as it contains consensus sequences present in proteins of the WASP family, including an actin monomer-binding domain and two acidic regions that interact with and activate the Arp2/3 complex, a host actin nucleator. [164][165][166][167][168][169][170][171] However, ActA lacks regulatory sequences that are present in WASP proteins that bind to Rho family GTPases. The central domain of ActA, a proline-rich repeat region, is required to recruit proteins of the Ena/ VASP protein family, which modulates bacterial speed and directionality.…”

Section: ©2 0 1 1 L a N D E S B I O S C I E N C E D O N O T D I S Tmentioning

confidence: 99%

The arsenal of virulence factors deployed byListeria monocytogenesto promote its cell infection cycle

et al. 2011

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“…In contrast, a listerial outer membrane protein, ActA, does not bind with Nck or N-WASP, but instead directly binds with Arp2/3, through the ActA region shared with WASP family proteins. This region mimics the activity of WASP, thereby creating actin tails attached to one pole of a bacterial cell (6,30,49,58). Thus, a signal through N-WASP or a WASP-related molecule is considered to be a common feature for the formation of actin-rich structures, although the bacterial elements triggering the formation differ in different bacteria.…”

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confidence: 99%

Enteropathogenic Escherichia coli , Shigella flexneri , and Listeria monocytogenes Recruit a Junctional Protein, Zonula Occludens-1, to Actin Tails and Pedestals

et al. 2007

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Enteropathogenic Escherichia coli, Shigella flexneri, and Listeria monocytogenes induce localized actin polymerization at the cytoplasmic face of the plasma membrane or within the host cytoplasm, creating unique actin-rich structures termed pedestals or actin tails. The process is known to be mediated by the actin-related protein 2 and 3 (Arp2/3) complex, which in these cases acts downstream of neural Wiskott-Aldrich syndrome protein (N-WASP) or of a listerial functional homolog of WASP family proteins. Here, we show that zonula occludens-1 (ZO-1), a protein in the tight junctions of polarized epithelial cells, is recruited to actin tails and pedestals. Immunocytochemical analysis revealed that ZO-1 was stained most in the distal part of the actin-rich structures, and the incorporation was mediated by the proline-rich region of the ZO-1 molecule. The direct clustering of membrane-targeted Nck, which is known to activate the N-WASP-Arp2/3 pathway, triggered the formation of the ZO-1-associated actin tails. The results suggest that the activation of the Arp2/3 complex downstream of N-WASP or a WASP-related molecule is a key to the formation of the particular actin-rich structures that bind with ZO-1. We propose that an analysis of the recruitment on a molecular basis will lead to an understanding of how ZO-1 recognizes a distinctive actin-rich structure under pathophysiological conditions.

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Listeria Protein ActA Mimics WASP Family Proteins: It Activates Filament Barbed End Branching by Arp2/3 Complex

Cited by 117 publications

References 48 publications

Phosphorylation of actin Tyr-53 inhibits filament nucleation and elongation and destabilizes filaments

Phosphorylation of actin Tyr-53 inhibits filament nucleation and elongation and destabilizes filaments

The arsenal of virulence factors deployed byListeria monocytogenesto promote its cell infection cycle

Enteropathogenic Escherichia coli , Shigella flexneri , and Listeria monocytogenes Recruit a Junctional Protein, Zonula Occludens-1, to Actin Tails and Pedestals

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