Sequence determination and mutational analysis of the lly locus of Legionella pneumophila

Wintermeyer, Eva; Flügel, M; Ott, Manfred; Steinert, Michael; Rdest, Ursula; Mann, Karlheinz; Häcker, J.

doi:10.1128/iai.62.3.1109-1117.1994

Cited by 47 publications

(43 citation statements)

References 63 publications

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“…One such gene that in£uences bacterial survival in the natural environment is the lly (legiolysin) locus. Mutagenesis of the lly gene of the L. pneumophila Philadelphia I derivative JR32 did not a¡ect intracellular replication in natural amoebal hosts or in U937 macrophage-like cells [3,4]. However, the Lly-negative mutant showed a markedly decreased resistance to light, indicating a contribution of the Lly protein to ecological adaption of Legionella [3].…”

Section: Introductionmentioning

confidence: 93%

“…pneumophila strains Philadelphia I JR 32 and Corby and plasmids used in this study are described elsewhere [3,4,6,7]. The transcomplementation of the lly-negative mutant JR 32-1 resulted in L. pneumophila JR32-1 (pEWB 34-114) [4]. For expression of the Lly protein in E. coli DH5K, the plasmids pEWL-1 and pEWL-113 were used [6].…”

Section: Bacterial Strains and Plasmidsmentioning

confidence: 99%

“…To analyse the complete DNA sequence and the function of the lly gene of strain Corby, the chromosomal lly gene was ampli¢ed using the sense primer 5P-GAATTCGTTAATGAATTGAATGG-3P and antisense primer 5P-TAGATCTCCTGGCTTATGC-3P, designed on the basis of the published sequence [4] using the software package Lasergene. The PCR product of 2653 bp was puri¢ed with the QIAquick puri¢cation kit (Qiagen, Hilden, Germany), ligated into vector pPCR-Script (Stratagene, Heidelberg, Germany) and transformed into E. coli Epicurian XL1-blue cells.…”

Section: Ampli¢cation and Cloning Of The Complete Lly Gene Of L Pneumentioning

confidence: 99%

“…To produce the anti-legiolysin mAb 16/1 for Western blotting of total bacterial extracts balb/c mice were immu-nised with the recombinant E. coli strain DH5K/pEWL-113 [4,9]. Plasmid pEWL-113 contained a 2.4-kb EcoRI-SmaI fragment of the lly locus [4].…”

Section: Western Blotting Analysis Of Legiolysin Proteins Using Mab 16/1mentioning

confidence: 99%

“…The lly gene encodes for a 39-kDa protein that confers haemolysis, browning of the culture medium, and £uorescence of recombinant Escherichia coli K-12 clones [6]. The nucleotide sequence of the lly gene contains an open reading frame that codes for 348 amino acid residues [4].…”

Section: Introductionmentioning

confidence: 99%

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The Lly protein is essential for p-hydroxyphenylpyruvate dioxygenase activity in Legionella pneumophila

Steinert

2001

FEMS Microbiology Letters

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The lly locus confers fluorescence, haemolysis, brown pigmentation and an increased resistance to light in Legionella pneumophila. In this study, we correlated the pigment production of two lly-positive L. pneumophila isolates and a recombinant lly-positive Escherichia coli strain with the presence of homogentisic acid (HGA) in the culture supernatant. The detection of HGA by high performance liquid chromatography and the data analysis of the deduced amino acid sequence of the lly gene indicate that the lly locus codes for a p-hydroxyphenylpyruvate dioxygenase (HPPD). This enzyme catalyses the transformation of p-hydroxyphenylpyruvate into HGA, which subsequently oxidises and polymerises into a melanin-like pigment. One open reading frame (ORF 1) in the lly region exhibited homologies with genes of Synechocystis sp., Petroselium crispum and Streptomyces mycarofaciens that code for methyltransferases. By screening a genomic library of L. pneumophila (serogroup 1) strain Corby with a monoclonal antibody against the legiolysin (lly), we identified two recombinant E. coli clones that did not produce the brown pigment and showed no haemolysis and fluorescence. DNA sequencing revealed that both clones contained 874 nucleotides of the N-terminal part of the lly gene. The recombinant strains expressed truncated legiolysin proteins of 39.5 and 35.7 kDa and did not produce HGA. Considering the highly conserved structure of legiolysin-like HPPD genes from other organisms, we suggest that the C-terminus of the legiolysin may be essential for the enzymatic activity that conferred pigmentation via HGA polymerisation, haemolysis and fluorescence. ß

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Section: Introductionmentioning

confidence: 93%

Section: Bacterial Strains and Plasmidsmentioning

confidence: 99%

Section: Ampli¢cation and Cloning Of The Complete Lly Gene Of L Pneumentioning

confidence: 99%

Section: Western Blotting Analysis Of Legiolysin Proteins Using Mab 16/1mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The Lly protein is essential for p-hydroxyphenylpyruvate dioxygenase activity in Legionella pneumophila

Steinert

2001

FEMS Microbiology Letters

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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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THE TETRAHYMENA HOMOLOG OF BACTERIAL AND MAMMALIAN 4‐HYDROXYPHENYLPYRUVATE DIOXYGENASES LOCALIZES TO MEMBRANES OF THE ENDOPLASMIC RETICULUM

Neve¹,

Tornehave²,

Corydon

et al. 1999

Cell Biology International

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The expression and intracellular localization of the Tetrahymena homolog of 4-hydroxyphenylpyruvate dioxygenase (HPPD) were investigated in wild-type Tetrahymena thermophila strain B1868 VII and the mutant strains IIG8, defective in food vacuole formation, MS-1, blocked in secretion of lysosomal enzymes, and SB 281, defective in mucocyst maturation. Immunoelectron microscopy and confocal laser scanning microscopy demonstrated that Tetrahymena HPPD primarily localized to membranes of the endoplasmic reticulum. In addition, Tetrahymena HPPD was detected in association with membranes of the Golgi apparatus, and transport vesicles in exponentially growing wild-type and mutant strains. In starved cells, Tetrahymena HPPD localized exclusively to membranes of small vesicles. Since no de novo synthesis of Tetrahymena HPPD takes place in cells starved for more than 30 min, these results suggest that there is a flow of Tetrahymena HPPD from the endoplasmic reticulum to small vesicles, possibly via the Golgi apparatus, and that Tetrahymena HPPD contains a signal for vesicle membrane retrieval or retention.

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Sequence determination and mutational analysis of the lly locus of Legionella pneumophila

Cited by 47 publications

References 63 publications

The Lly protein is essential for p-hydroxyphenylpyruvate dioxygenase activity in Legionella pneumophila

The Lly protein is essential for p-hydroxyphenylpyruvate dioxygenase activity in Legionella pneumophila

The life stage‐specific pathometabolism of Legionella pneumophila

THE TETRAHYMENA HOMOLOG OF BACTERIAL AND MAMMALIAN 4‐HYDROXYPHENYLPYRUVATE DIOXYGENASES LOCALIZES TO MEMBRANES OF THE ENDOPLASMIC RETICULUM

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