Amino Acid Uptake and Metabolism of Legionella pneumophila Hosted by Acanthamoeba castellanii

Schunder, Eva; Gillmaier, Nadine; Kutzner, Erika; Herrmann, Vroni; Lautner, Monika; Heuner, Klaus; Eisenreich, Wolfgang

doi:10.1074/jbc.m114.570085

Cited by 51 publications

(74 citation statements)

References 74 publications

(125 reference statements)

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“…The following isogenic mutant strains were used: ⌬keto (lpp1788; acetyl-CoA acetyltransferase, ␤-ketothiolase) (14), ⌬zwf (lpp0483; glucose-6-phosphate-dehydrogenase) (19), ⌬gam (lpp0489; glucoamylase) (34), ⌬lpp0931-33, ⌬keto/⌬lpp0931-33, and ⌬lpp0650 mutant strains (this work; see below). Escherichia coli DH5␣, serving as host for amplification of recombinant plasmid DNA, was grown in lysogeny broth (LB) or on LB agar (37,38 L. pneumophila was grown in ACES-buffered yeast extract (AYE) broth consisting of 10 g of ACES, 10 g of yeast extract, 0.4 g of L-Cys, and 0.25 g of ferric pyrophosphate/liter (adjusted to pH 6.8 with 3 M KOH and sterile filtrated) at 37°C with agitation at 250 rpm or on buffered charcoal-yeast extract agar for 3 days at 37°C.…”

Section: Methodsmentioning

confidence: 99%

“…For chromosomal recombination, the construct was amplified by PCR. Natural transformation of L. pneumophila Paris was done as described before with modification (14,41). The ⌬keto/⌬lpp0931-33 double mutant was constructed by using the ⌬keto strain as the acceptor strain for natural transformation using the PCR product of the lpp0931-33-Gm R cassette construct (see above).…”

Section: Methodsmentioning

confidence: 99%

“…encodes one of the four putative PHB polymerases in L. pneumophila, and lpp1788 putatively encodes the 3-ketothiolase reaction (14,32) (Fig. 1).…”

Section: Construction and Growth Characterization Of L Pneumophila Mmentioning

confidence: 99%

“…1). In L. pneumophila Paris, three putative 3-ketothiolases (Lpp1788, Lpp1555, and Lpp1307), three putative acetoacetyl-CoA reductases (Lpp0620, Lpp0621, and Lpp2322), and four putative PHB synthases (Lpp0650, Lpp2038, Lpp2214, and Lpp2323) can be assigned on the basis of sequence homologies (13,14). However, a functional assignment of these proteins is missing.…”

mentioning

confidence: 99%

“…1). Amino acids also play an important role as nutrients during growth within host cells (14,20,(27)(28)(29).…”

mentioning

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: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…encodes one of the four putative PHB polymerases in L. pneumophila, and lpp1788 putatively encodes the 3-ketothiolase reaction (14,32) (Fig. 1).…”

Section: Construction and Growth Characterization Of L Pneumophila Mmentioning

confidence: 99%

mentioning

confidence: 99%

“…1). Amino acids also play an important role as nutrients during growth within host cells (14,20,(27)(28)(29).…”

mentioning

confidence: 99%

See 3 more Smart Citations

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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The life stage‐specific pathometabolism of Legionella pneumophila

Eisenreich

Heuner

2016

FEBS Letters

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The genus Legionella belongs to Gram-negative bacteria found ubiquitously in aquatic habitats, where it grows in natural biofilms and replicates intracellularly in various protozoa (amoebae, ciliates). L. pneumophila is known as the causative agent of Legionnaires' disease, since it is also able to replicate in human alveolar macrophages, finally leading to inflammation of the lung and pneumonia. To withstand the degradation by its host cells, a Legionellacontaining vacuole (LCV) is established for intracellular replication, and numerous effector proteins are secreted into the host cytosol using a type four B secretion system (T4BSS). During intracellular replication, Legionella has a biphasic developmental cycle that alternates between a replicative and a transmissive form. New knowledge about the host-adapted and life stagedependent metabolism of intracellular L. pneumophila revealed a bipartite metabolic network with life stage-specific usages of amino acids (e.g. serine), carbohydrates (e.g. glucose) and glycerol as major substrates. These metabolic features are associated with the differentiation of the intracellular bacteria, and thus have an important impact on the virulence of L. pneumophila.

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Crosstalk between Metabolism and Virulence ofLegionella pneumophila

Heuner

Eisenreich

2016

Host – Pathogen Interaction

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Amino Acid Uptake and Metabolism of Legionella pneumophila Hosted by Acanthamoeba castellanii

Cited by 51 publications

References 74 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 life stage‐specific pathometabolism of Legionella pneumophila

Crosstalk between Metabolism and Virulence ofLegionella pneumophila

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