Sequence determination and mutational analysis of the lly locus of Legionella pneumophila.
The lly (legiolysin) locus codes for a 39-kDa protein which confers hemolysis, pigment production, and fluorescence on recombinant Escherichia coli K-12 clones carrying the lly gene. The nucleotide sequences of the lly genes from two Legionella pneumophila isolates were determined. The lly loci exhibited identical nucleotide sequences. They contained open reading frames of 348 amino acid residues, encoding proteins with a deduced molecular mass of 38.9 kDa. N-terminal amino acid sequencing further confirmed that the Lly protein corresponds to the open reading frame sequenced. The amino acid sequence of the Lly protein exhibits a high degree of homology with the sequences of the MelA protein responsible for melanin production in the freshwater bacterium Shewanella colwelliana and the 4-hydroxyphenylpyruvate dioxygenase of Pseudomonas spp. 4-Hydroxyphenylpyruvate dioxygenase is involved in the degradation of aromatic amino acids in various organisms. An Lly-negative mutant of L. pneumophila Philadelphia I derivative JR32 and an Lly-positive transcomplementant were constructed. The Lly-negative mutant lost the ability to produce brown pigment and to confer fluorescence but retained hemolysis. Introduction of a plasmid carrying the lly locus restored pigment production and fluorescence. Intracellular survival of L. pneumophila in U937 macrophage-like cells and in Acanthamoeba castellanii was not affected by mutagenization of the lly locus.
Wintermeyer
E
E
Institut für Molekulare Infektionsbiologie, Universität Würzburg, Germany.
Flügel
M
M
Ott
M
M
Steinert
M
M
Rdest
U
U
Mann
K H
KH
Hacker
J
J
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D002874:Chromosome Mapping; D004252:DNA Mutational Analysis; D005798:Genes, Bacterial; D006461:Hemolysis; D006801:Humans; D016952:Legionella pneumophila / Q000235:genetics* / Q000254:growth & development; D008969:Molecular Sequence Data; D014407:Tumor Cells, Cultured
D001426:Bacterial Proteins; C094283:Lly protein, Legionella pneumophila