Listeria monocytogenes ActA protein interacts with phosphatidylinositol 4,5-bisphosphate in vitro

Steffen, Pierre; Schafer, Dorothy A.; David, Violaine; Gouin, Edith; Cooper, John A.; Cossart, Pascale

doi:10.1002/(sici)1097-0169(200001)45:1<58::aid-cm6>3.0.co;2-y

Cited by 28 publications

(19 citation statements)

References 34 publications

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“…Therefore, it is possible that PlcA acts in a vacuole to counteract autophagy (25) and/or acts globally to impact both autophagy (26) and/or other functions. For instance, PlcA activity might affect actin-based motility, since PI(3,5)P 2 and PI(3,4,5)P 3 bind to ActA (56,57).…”

Section: Discussionmentioning

confidence: 99%

Avoidance of Autophagy Mediated by PlcA or ActA Is Required for Listeria monocytogenes Growth in Macrophages

Mitchell

Huang

et al. 2015

Infect Immun

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Listeria monocytogenes is a facultative intracellular pathogen that escapes from phagosomes and grows in the cytosol of infected host cells. Most of the determinants that govern its intracellular life cycle are controlled by the transcription factor PrfA, including the pore-forming cytolysin listeriolysin O (LLO), two phospholipases C (PlcA and PlcB), and ActA. We constructed a strain that lacked PrfA but expressed LLO from a PrfA-independent promoter, thereby allowing the bacteria to gain access to the host cytosol. This strain did not grow efficiently in wild-type macrophages but grew normally in macrophages that lacked ATG5, a component of the autophagy LC3 conjugation system. This strain colocalized more with the autophagy marker LC3 (42% ؎ 7%) at 2 h postinfection, which constituted a 5-fold increase over the colocalization exhibited by the wild-type strain (8% ؎ 6%). While mutants lacking the PrfA-dependent virulence factor PlcA, PlcB, or ActA grew normally, a double mutant lacking both PlcA and ActA failed to grow in wild-type macrophages and colocalized more with LC3 (38% ؎ 5%). Coexpression of LLO and PlcA in a PrfA-negative strain was sufficient to restore intracellular growth and decrease the colocalization of the bacteria with LC3. In a cell-free assay, purified PlcA protein blocked LC3 lipidation, a key step in early autophagosome biogenesis, presumably by preventing the formation of phosphatidylinositol 3-phosphate (PI3P). The results of this study showed that avoidance of autophagy by L. monocytogenes primarily involves PlcA and ActA and that either one of these factors must be present for L. monocytogenes growth in macrophages.

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Section: Discussionmentioning

confidence: 99%

Avoidance of Autophagy Mediated by PlcA or ActA Is Required for Listeria monocytogenes Growth in Macrophages

Mitchell

Huang

et al. 2015

Infect Immun

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“…Rabbit polyclonal antibodies to ActA (used at 1:500) were generated as previously described. 20 All secondary antibodies used were AlexaFluor conjugates (used at 1:200) from Molecular Probes. DAPI (Molecular Probes) was used according to the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

Listeria monocytogenesEvades Killing by Autophagy During Colonization of Host Cells

Birmingham

Canadien²,

Gouin³

et al. 2007

Autophagy

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226

206

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Previously published online as an Autophagy E-publication: http://www.landesbioscience.com/journals/autophagy/abstract.php?id=4450 KEY worDsListeria monocytogenes, bacterial invasion, autophagy, innate immunity, lysosome, macrophage. ABBrEViATions ABsTrACTListeria monocytogenes is an intracellular pathogen that is able to colonize the cytosol of macrophages. Here we examined the interaction of this pathogen with autophagy, a host cytosolic degradative pathway that constitutes an important component of innate immunity towards microbial invaders. L. monocytogenes infection induced activation of the autophagy system in macrophages. At 1 h post infection (p.i.), a population of intracellular bacteria (~37%) colocalized with the autophagy marker LC3. These bacteria were within vacuoles and were targeted by autophagy in an LLO-dependent manner. At later stages in infection (by 4 h p.i.), the majority of L. monocytogenes escaped into the cytosol and rapidly replicated. At these times, less than 10% of intracellular bacteria colocalized with LC3. We found that ActA expression was sufficient to prevent autophagy of bacteria in the cytosol of macrophages. Surprisingly, ActA expression was not strictly necessary, indicating that other virulence factors were involved. Accordingly, we also found a role for the bacterial phospholipases, PI-PLC and PC-PLC, in autophagy evasion, as bacteria lacking phospholipase expression were targeted by autophagy at later times in infection. Together, our results demonstrate that L. monocytogenes utilizes multiple mechanisms to avoid destruction by the autophagy system during colonization of macrophages.

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“…The data are not convincing, however, since ActA binds the D-3 phosphoinositides better than PI(4,5)P 2 (28), and the D-3 phosphoinositides that have been directly tested do not inhibit capping protein activity (152). Moreover, ActA binding diminishes the PI(4,5)P 2 -mediated inhibition of capping protein activity (166).…”

Section: Interaction Of Phosphoinositides With Actamentioning

confidence: 97%

“…Phosphoinositides also bind ActA in vitro (28,166). Binding of phosphoinositides to ActA induces a conformational change in the amino-terminal portion of the protein (amino acids 29 to 267) (28).…”

Section: Interaction Of Phosphoinositides With Actamentioning

confidence: 99%

Actin-Based Motility of Intracellular Microbial Pathogens

Goldberg

2001

Microbiol Mol Biol Rev

199

188

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A diverse group of intracellular microorganisms, including Listeria monocytogenes, Shigella spp., Rickettsia spp., and vaccinia virus, utilize actin-based motility to move within and spread between mammalian host cells. These organisms have in common a pathogenic life cycle that involves a stage within the cytoplasm of mammalian host cells. Within the cytoplasm of host cells, these organisms activate components of the cellular actin assembly machinery to induce the formation of actin tails on the microbial surface. The assembly of these actin tails provides force that propels the organisms through the cell cytoplasm to the cell periphery or into adjacent cells. Each of these organisms utilizes preexisting mammalian pathways of actin rearrangement to induce its own actin-based motility. Particularly remarkable is that while all of these microbes use the same or overlapping pathways, each intercepts the pathway at a different step. In addition, the microbial molecules involved are each distinctly different from the others. Taken together, these observations suggest that each of these microbes separately and convergently evolved a mechanism to utilize the cellular actin assembly machinery. The current understanding of the molecular mechanisms of microbial actin-based motility is the subject of this review

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Listeria monocytogenes ActA protein interacts with phosphatidylinositol 4,5-bisphosphate in vitro

Cited by 28 publications

References 34 publications

Avoidance of Autophagy Mediated by PlcA or ActA Is Required for Listeria monocytogenes Growth in Macrophages

Avoidance of Autophagy Mediated by PlcA or ActA Is Required for Listeria monocytogenes Growth in Macrophages

Listeria monocytogenesEvades Killing by Autophagy During Colonization of Host Cells

Actin-Based Motility of Intracellular Microbial Pathogens

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