GamA is a eukaryotic-like glucoamylase responsible for glycogen- and starch-degrading activity of Legionella pneumophila

Herrmann, Vroni; Eidner, Anja; Rydzewski, Kerstin; Blädel, Inga; Jules, Matthieu; Buchrieser, Carmen; Eisenreich, Wolfgang; Heuner, Klaus

doi:10.1016/j.ijmm.2010.08.016

Cited by 35 publications

(37 citation statements)

References 39 publications

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“…In addition, it also supports the published role of the ED pathway for the life cycle of L. pneumophila (19,33). The gamA gene encodes a glucoamylase responsible for the glycogen-degrading activity of L. pneumophila Paris, but the inactivation of gamA had no effect on intracellular replication in A. castellanii (34). As expected, the amount of PHB of the ⌬gam mutant strain was similar to that of the WT strain (Fig.…”

Section: Construction and Growth Characterization Of L Pneumophila Msupporting

confidence: 83%

“…It was also reported that the ED pathway is necessary during the intracellular life cycle of L. pneumophila (33). Indeed, host cell glycogen could be degraded to glucose by the action of the bacterial glucoamylase GamA (19,34). Further supporting the role of glucose as a nutrient for intracellular L. pneumophila, glucose uptake was found to be increased during the late phases of growth (33), and Legionella species-specific differences in their usages of glucose and serine as carbon substrates were suggested recently (35,36).…”

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confidence: 99%

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Growth-related Metabolism of the Carbon Storage Poly-3-hydroxybutyrate in Legionella pneumophila

Gillmaier

Schunder

Kutzner

et al. 2016

Journal of Biological Chemistry

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Legionella pneumophila, the causative agent of Legionnaires disease, has a biphasic life cycle with a switch from a replicative to a transmissive phenotype. During the replicative phase, the bacteria grow within host cells in Legionella-containing vacuoles. During the transmissive phenotype and the postexponential (PE) growth phase, the pathogens express virulence factors, become flagellated, and leave the Legionella-containing vacuoles. Using 13 C labeling experiments, we now show that, under in vitro conditions, serine is mainly metabolized during the replicative phase for the biosynthesis of some amino acids and for energy generation. During the PE phase, these carbon fluxes are reduced, and glucose also serves as an additional carbon substrate to feed the biosynthesis of poly-3-hydroxybuyrate (PHB), an essential carbon source for transmissive L. pneumophila. Whole-cell FTIR analysis and comparative isotopologue profiling further reveal that a putative 3-ketothiolase (Lpp1788) and a PHB polymerase (Lpp0650), but not enzymes of the crotonylCoA pathway (Lpp0931-0933) are involved in PHB metabolism during the PE phase. However, the data also reflect that additional bypassing reactions for PHB synthesis exist in agreement with in vivo competition assays using Acanthamoeba castellannii or human macrophage-like U937 cells as host cells. The data suggest that substrate usage and PHB metabolism are coordinated during the life cycle of the pathogen.

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Section: Construction and Growth Characterization Of L Pneumophila Msupporting

confidence: 83%

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confidence: 99%

Growth-related Metabolism of the Carbon Storage Poly-3-hydroxybutyrate in Legionella pneumophila

Gillmaier

Schunder

Kutzner

et al. 2016

Journal of Biological Chemistry

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“…Cell-free supernatants collected from late-exponential BYE cultures were assayed for protease activity as measured by azocasein hydrolysis, for phosphatase activity as measured by the release of p-nitrophenol from p-nitrophenol phosphate, for lipase activity as measured by the release of p-nitrophenol from p-nitrophenol caprylate, for chitinase activity as measured by the release of p-nitrophenol from the chromogenic substrate p-nitrophenol-[GlcNAc] 3 , and for leucine and lysine aminopeptidase activities as measured by the release of p-nitroanilide (pNI) from L-leucine p-NI and L-lysine p-NI, respectively (Aragon et al, 2000(Aragon et al, , 2001DebRoy et al, 2006b;Rossier et al, 2008). Starch-degrading activity was monitored as previously described (Herrmann et al, 2011;Tyson et al, 2013). Swimming motility was determined by wet mount, and sliding motility was assessed by examining bacteria spotted onto BCYE containing only 0.5 % agar (Stewart et al, 2011).…”

Section: Methodsmentioning

confidence: 99%

The novel Legionella pneumophila type II secretion substrate NttC contributes to infection of amoebae Hartmannella vermiformis and Willaertia magna

2014

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The type II protein secretion (T2S) system of Legionella pneumophila secretes over 25 proteins, including novel proteins that have no similarity to proteins of known function. T2S is also critical for the ability of L. pneumophila to grow within its natural amoebal hosts, including Acanthamoeba castellanii, Hartmannella vermiformis and Naegleria lovaniensis. Thus, T2S has an important role in the natural history of legionnaires' disease. Our previous work demonstrated that the novel T2S substrate NttA promotes intracellular infection of A. castellanii, whereas the secreted RNase SrnA, acyltransferase PlaC, and metalloprotease ProA all promote infection of H. vermiformis and N. lovaniensis. In this study, we determined that another novel T2S substrate that is specific to Legionella, designated NttC, is unique in being required for intracellular infection of H. vermiformis but not for infection of N. lovaniensis or A. castellanii. Expanding our repertoire of amoebal hosts, we determined that Willaertia magna is susceptible to infection by L. pneumophila strains 130b, Philadelphia-1 and Paris. Furthermore, T2S and, more specifically, NttA, NttC and PlaC were required for infection of W. magna. Taken together, these data demonstrate that the T2S system of L. pneumophila is critical for infection of at least four types of aquatic amoebae and that the importance of the individual T2S substrates varies in a host cell-specific fashion. Finally, it is now clear that novel T2S-dependent proteins that are specific to the genus Legionella are particularly important for L. pneumophila infection of key, environmental hosts.

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“…This may explain the inability of L. oakridgensis to degrade starch and glycogen. GamA was shown to be responsible for glycogen-degrading activity in L. pneumophila (Herrmann et al, 2011). In addition, our isotopologue profiling experiments demonstrated that L. oakridgensis, in contrast to L. pneumophila, is not able to use supplied glucose as a carbon source for de novo synthesis of amino acids or PHB.…”

Section: Major Pathways Of the Metabolism Carbohydrate Utilization Amentioning

confidence: 77%

“…Alpha-amylase activity plates contained 0.1% starch or glycogen and cellulose-activity plates were supplemented with 0.1% carboxymethyl cellulose (CMC, Sigma-Aldrich, St.-Leon-Roth, Germany). Experiments were carried out as described previously (Herrmann et al, 2011). Protein degrading activity was visualized on agar containing 0.9% casein (C8654, Sigma-Aldrich, St.-Leon-Roth, Germany) and 0.14% starch (Roth, Karlsruhe, Germany).…”

Section: Phenotypic Assaysmentioning

confidence: 99%

Legionella oakridgensis ATCC 33761 genome sequence and phenotypic characterization reveals its replication capacity in amoebae

Brzuszkiewicz

Schulz

Rydzewski

et al. 2013

International Journal of Medical Microbiology

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Legionella oakridgensis is able to cause Legionnaires` disease, but is less virulent compared to L. pneumophila strains and very rarely associated with human disease. L. oakridgensis is the only species of the family legionellae which is able to grow on media without additional cysteine. In contrast to earlier publications, we found that L. oakridgensis is able to multiply in amoebae. We sequenced the genome of L. oakridgensis type strain OR-10 (ATCC 33761).The genome is smaller than the other yet sequenced Legionella genomes and has a higher G+C-content of 40.9%. L. oakridgensis lacks a flagellum and it also lacks all genes of the flagellar regulon except of the alternative sigma-28 factor FliA and the anti-sigma-28 factor

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GamA is a eukaryotic-like glucoamylase responsible for glycogen- and starch-degrading activity of Legionella pneumophila

Cited by 35 publications

References 39 publications

Growth-related Metabolism of the Carbon Storage Poly-3-hydroxybutyrate in Legionella pneumophila

Growth-related Metabolism of the Carbon Storage Poly-3-hydroxybutyrate in Legionella pneumophila

The novel Legionella pneumophila type II secretion substrate NttC contributes to infection of amoebae Hartmannella vermiformis and Willaertia magna

Legionella oakridgensis ATCC 33761 genome sequence and phenotypic characterization reveals its replication capacity in amoebae

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