Genetic identification of chemotactic transducers for amino acids in Pseudomonas aeruginosa.
Two chemotactic transducer genes (termed pctB and pctC) and an open reading frame (orf1) were found in the pctA-flanking region which was previously identified as a chemotactic transducer gene in Pseudomonas aeruginosa. The pctB and pctC genes encode predicted polypeptides of 629 and 632 amino acids, respectively. Overall, PctB and PctC had 81 and 75% amino acid identities with PctA, respectively. A null mutant strain PCT2, which contained a deletion in the entire pctC, orf1, pctA and pctB genes, did not show chemotaxis towards all 20 commonly occurring L-amino acids. This mutant strain also failed to respond to amino acid catabolites (cadaverine, 4-aminobutyrate and putrescine) that are strong attractants for the wild-type strain PAO1. To study the role of each gene product in L-amino acid taxis, plasmids harbouring the pctC, orf1, pctA, or pctB genes were constructed and introduced into strain PCT2 by transformation. The orf1 gene did not complement the defect in chemotaxis of strain PCT2. The pctA gene restored the ability of strain PCT2 to respond to 18 L-amino acids, suggesting that PctA plays a major role in detecting L-amino acids in P. aeruginosa. The pctB and pctC genes complemented the defect in chemotaxis to only seven (Ala, Arg, Glu, Lys, Met, Tyr, Gln) and two (His, Pro) L-amino acids, respectively.
Taguchi
Kazunori
K
Department of Fermentation Technology, Hiroshima University, Higashi-Hiroshima, Hiroshima 739, Japan.
Fukutomi
Hiroyuki
H
Department of Fermentation Technology, Hiroshima University, Higashi-Hiroshima, Hiroshima 739, Japan.
Kuroda
Akio
A
Department of Fermentation Technology, Hiroshima University, Higashi-Hiroshima, Hiroshima 739, Japan.
Kato
Junichi
J
Department of Fermentation Technology, Hiroshima University, Higashi-Hiroshima, Hiroshima 739, Japan.
Ohtake
Hisao
H
Department of Fermentation Technology, Hiroshima University, Higashi-Hiroshima, Hiroshima 739, Japan.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics; D002633:Chemotaxis / Q000187:drug effects / Q000235:genetics*; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D005810:Multigene Family; D016254:Mutagenesis, Insertional; D009154:Mutation; D016366:Open Reading Frames; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000235:genetics* / Q000502:physiology*; D015183:Restriction Mapping; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D014170:Transformation, Genetic
D000596:Amino Acids; D001426:Bacterial Proteins; C110124:PctA protein, Pseudomonas aeruginosa; C110122:PctB protein, Pseudomonas aeruginosa; C110123:PctC protein, Pseudomonas aeruginosa