Identification and functional characterization of a Xenorhabdus nematophila oligopeptide permease

• Published in: Applied and Environmental Microbiology 2004-09, Vol.70 (9), p.5621-5627
• Main Authors: Orchard, S.S, Goodrich-Blair, H
• Format: Article
• Language: English
• Subjects: ABC transporters; Bacteria; bacterial proteins; Bacterial Proteins - genetics; Bacterial Proteins - isolation & purification; Bacterial Proteins - metabolism; Bacteriology; binding proteins; Biological and medical sciences; Cloning, Molecular; Conjugation, Genetic - genetics; entomopathogenic bacteria; Enzymes; Enzymology and Protein Engineering; Escherichia coli - enzymology; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; genes; hemolymph; Insects; leuA gene; leuB gene; loci; Manduca sexta; Membrane Transport Proteins - genetics; Membrane Transport Proteins - isolation & purification; Membrane Transport Proteins - metabolism; Metabolism. Enzymes; microbial growth; Microbiology; Molecular Sequence Data; mutants; nucleotide sequences; oligopeptide-binding proteins; oligopeptides; Operon - genetics; opp genes; opp loci; Pathogens; Peptides; Plasmids - genetics; Steinernema carpocapsae; Xenorhabdus - enzymology; Xenorhabdus - genetics; Xenorhabdus nematophila
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/AEM.70.9.5621-5627.2004

The bacterium Xenorhabdus nematophila is a mutualist of Steinernema carpocapsae nematodes and a pathogen of insects. Presently, it is not known what nutrients the bacterium uses to thrive in these host environments. In other symbiotic bacteria, oligopeptide permeases have been shown to be important in host interactions, and we therefore sought to determine if oligopeptide uptake is essential for growth or symbiotic functions of X. nematophila in laboratory or host environments. We identified an X. nematophila oligopeptide permease (opp) operon of two sequential oppA genes, predicted to encode oligopeptide-binding proteins, and putative permease-encoding genes oppB, oppC, oppD, and oppF. Peptide-feeding studies indicated that this opp operon encodes a functional oligopeptide permease. We constructed strains with mutations in oppA1, oppA2, or oppB and examined the ability of each mutant strain to grow in a peptide-rich laboratory medium and to interact with the two hosts. We found that the opp mutant strains had altered growth phenotypes in the laboratory medium and in hemolymph isolated from larval insects. However, the opp mutant strains were capable of initiating and maintaining both mutualistic and pathogenic host interactions. These data demonstrate that the opp genes allow X. nematophila to utilize peptides as a nutrient source but that this function is not essential for the existence of X. nematophila in either of its host niches. To our knowledge, this study represents the first experimental analysis of the role of oligopeptide transport in mediating a mutualistic invertebrate-bacterium interaction.