Requirement for Formin-Induced Actin Polymerization during Spread of
            <i>Shigella flexneri</i>

Heindl, Jason E.; Saran, Indrani; Yi, Chae-ryun; Lesser, Cammie F.; Goldberg, Marcia B.

doi:10.1128/iai.00252-09

Cited by 56 publications

(62 citation statements)

References 50 publications

(56 reference statements)

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“…The speed of bacteria undergoing motility (Ͼ0.01 m/s) for at least 60 consecutive seconds was scored, and the period of sustained movement was recorded. (29), but various nonintestinal cell lines have been shown to support S. flexneri infection in tissue culture systems (17,23,25). Thus, most studies on the cellular determinants supporting S. flexneri actin-based motility have been conducted in nonintestinal cell lines.…”

Section: Methodsmentioning

confidence: 99%

“…However, another report showed that S. flexneri actin tails remained largely unchanged when N-WASP phosphodefective or phosphomimic mutant proteins were expressed (24). The vast majority of S. flexneri actin tail formation and intercellular motility studies have been conducted in nonintestinal cell lines such as HeLa cells and mouse fibroblasts (17,23,25). We sought to characterize the mechanism of actin tail formation in a relevant system for S. flexneri infection.…”

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confidence: 99%

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Bruton's Tyrosine Kinase Regulates Shigella flexneri Dissemination in HT-29 Intestinal Cells

2013

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Shigella flexneri is a Gram-negative intracellular pathogen that infects the intestinal epithelium and utilizes actin-based motility to spread from cell to cell. S. flexneri actin-based motility has been characterized in various cell lines, but studies in intestinal cells are limited. Here we characterized S. flexneri actin-based motility in HT-29 intestinal cells. In agreement with studies conducted in various cell lines, we showed that S. flexneri relies on neural Wiskott-Aldrich Syndrome protein (N-WASP) in HT-29 cells. We tested the potential role of various tyrosine kinases involved in N-WASP activation and uncovered a previously unappreciated role for Bruton's tyrosine kinase (Btk) in actin tail formation in intestinal cells. We showed that Btk depletion led to a decrease in N-WASP phosphorylation which affected N-WASP recruitment to the bacterial surface, decreased the number of bacteria displaying actin-based motility, and ultimately affected the efficiency of spread from cell to cell. Finally, we showed that the levels of N-WASP phosphorylation and Btk expression were increased in response to infection, which suggests that S. flexneri infection not only triggers the production of proinflammatory factors as previously described but also manipulates cellular processes required for dissemination in intestinal cells.

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

Bruton's Tyrosine Kinase Regulates Shigella flexneri Dissemination in HT-29 Intestinal Cells

2013

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“…In addition to the ARP2/3-dependent assembly machinery and the AIP1/cofilin-dependent disassembly machinery (23), several studies have revealed the importance of various cellular components in bacterial dissemination. These include the cell-cell adhesion protein E-cadherin (24), the gap junction protein connexin 26 (25), the myosin light chain kinase and its target myosin II (26,27), myosin 10 (28), the membrane-cytoskeleton linker ezrin (29), the dynamin binding protein Tuba (30), and actin nucleators of the formin family (31,32). Recent studies have also revealed the importance of cellular signaling in bacterial dissemination (33)(34)(35)(36).…”

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confidence: 99%

The Class II Phosphatidylinositol 3-Phosphate Kinase PIK3C2A Promotes Shigella flexneri Dissemination through Formation of Vacuole-Like Protrusions

Dragoi¹,

Agaisse²

2015

Infect Immun

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Intracellular pathogens such as Shigella flexneri and Listeria monocytogenes achieve dissemination in the intestinal epithelium by displaying actin-based motility in the cytosol of infected cells. As they reach the cell periphery, motile bacteria form plasma membrane protrusions that resolve into vacuoles in adjacent cells, through a poorly understood mechanism. Here, we report on the role of the class II phosphatidylinositol 3-phosphate kinase PIK3C2A in S. flexneri dissemination. Time-lapse microscopy revealed that PIK3C2A was required for the resolution of protrusions into vacuoles through the formation of an intermediate membrane-bound compartment that we refer to as a vacuole-like protrusion (VLP). Genetic rescue of PIK3C2A depletion with RNA interference (RNAi)-resistant cDNA constructs demonstrated that VLP formation required the activity of PIK3C2A in primary infected cells. PIK3C2A expression was required for production of phosphatidylinositol 3-phosphate [PtdIns(3)P] at the plasma membrane surrounding protrusions. PtdIns(3)P production was not observed in the protrusions formed by L. monocytogenes, whose dissemination did not rely on PIK3C2A. PIK3C2A-mediated PtdIns(3)P production in S. flexneri protrusions was regulated by host cell tyrosine kinase signaling and relied on the integrity of the S. flexneri type 3 secretion system (T3SS). We suggest a model of S. flexneri dissemination in which the formation of VLPs is mediated by the PIK3C2A-dependent production of the signaling lipid PtdIns(3)P in the protrusion membrane, which relies on the T3SS-dependent activation of tyrosine kinase signaling in protrusions. Shigella flexneri and Listeria monocytogenes are food-borne pathogens that display the ability to invade nonphagocytic cells, such as epithelial cells, and spread from primary infected cells to adjacent cells (1)(2)(3)(4). This dissemination process is supported by actin-based motility in primary infected cells (5, 6), which leads to the formation of membrane protrusions that project into adjacent cells, as motile bacteria reach the cell periphery (7,8). The resolution of protrusions into vacuoles from which the pathogen escapes allows the bacteria to gain access to the cytosolic compartment of adjacent cells, thereby achieving cell-to-cell spread (7,8). The mechanisms supporting L. monocytogenes and S. flexneri cytosolic motility are fairly well understood (9). Both pathogens achieve actin-based motility by recruiting to their surface a major nucleator of actin polymerization in eukaryotic cells, the ARP2/3 complex (10, 11). The expansion of the actin network formed at the bacterial surface generates forces that propel the bacterium throughout the cytosolic compartment (5, 6). In cells, the activity of the ARP2/3 complex is regulated by nucleationpromoting factors of the N-WASP/WAVE family (12, 13). S. flexneri engages the ARP2/3 complex through expression of IcsA (2, 14), a bacterial adaptor that recruits and activates N-WASP (15,16). L. monocytogenes does not engage the ARP2/3 complex through...

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“…Y. enterocolitica Yop proteins are known to specifically deactivate Rac1, Cdc42 and RhoA, while the role of Yops in counteracting Arf GTPases has not yet been reported (62). Induction of actin polymerization has been observed in other invasive pathogens, including L. monocytogenes (63), Shigella flexneri (64)(65)(66), and vaccinia virus (67,68). Likewise, L. monocytogenes uses the InlA and/or InlB surface protein to perform a "zippering" type of uptake using the same pathway (50)(51)(52).…”

Section: Discussionmentioning

confidence: 99%

Yersinia enterocolitica Inhibits Salmonella enterica Serovar Typhimurium and Listeria monocytogenes Cellular Uptake

et al. 2014

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bYersinia enterocolitica biovar 1B employs two type three secretion systems (T3SS), Ysa and Ysc, which inject effector proteins into macrophages to prevent phagocytosis. Conversely, Salmonella enterica serovar Typhimurium uses a T3SS encoded by Salmonella pathogenicity island 1 (SPI1) to actively invade cells that are normally nonphagocytic and a second T3SS encoded by SPI2 to survive within macrophages. Given the distinctly different outcomes that occur with regard to host cell uptake of S. Typhimurium and Y. enterocolitica, we investigated how each pathogen influences the internalization outcome of the other. Y. enterocolitica reduces S. Typhimurium invasion of HeLa and Caco-2 cells to a level similar to that observed using an S. Typhimurium SPI1 mutant alone. However, Y. enterocolitica had no effect on S. Typhimurium uptake by J774.1 or RAW264.7 macrophage-like cells. Y. enterocolitica was also able to inhibit the invasion of epithelial and macrophage-like cells by Listeria monocytogenes. Y. enterocolitica mutants lacking either the Ysa or Ysc T3SS were partially defective, while double mutants were completely defective, in blocking S. Typhimurium uptake by epithelial cells. S. Typhimurium encodes a LuxR homolog, SdiA, which detects N-acylhomoserine lactones (AHLs) produced by Y. enterocolitica and upregulates the expression of an invasin (Rck) and a putative T3SS effector (SrgE). Two different methods of constitutively activating the S. Typhimurium SdiA regulon failed to reverse the uptake blockade imposed by Y. enterocolitica.

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Requirement for Formin-Induced Actin Polymerization during Spread of Shigella flexneri

Cited by 56 publications

References 50 publications

Bruton's Tyrosine Kinase Regulates Shigella flexneri Dissemination in HT-29 Intestinal Cells

Bruton's Tyrosine Kinase Regulates Shigella flexneri Dissemination in HT-29 Intestinal Cells

The Class II Phosphatidylinositol 3-Phosphate Kinase PIK3C2A Promotes Shigella flexneri Dissemination through Formation of Vacuole-Like Protrusions

Yersinia enterocolitica Inhibits Salmonella enterica Serovar Typhimurium and Listeria monocytogenes Cellular Uptake

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