Intracellular receptor-type transcription factor, LasR, contains a highly conserved amphipathic region which precedes the putative helix-turn-helix DNA binding motif.
We have cloned and sequenced the lasR gene, which is involved in the transcriptional activation of several pathogenic factors, from Pseudomonas aeruginosa IFO3455 and PA103. These clones were predicted to be an open reading frame of 239 amino acids as reported for the PAO1 strain. There is only a single base change resulting in an amino acid exchange from M145 (PAO1) to I (IFO3455). PA103 DNA differs with PAO1 DNA in two bases resulting in only a single amino acid substitution from R179 to W. When the IFO3455 LasR was expressed in a PA103 strain which is known to be LasR defective, proteinase gene activation was detected, however, when PA103 LasR was expressed, no enhancement was measurable. From these results, it appears that the amino acid substitution of R179 to W inactivated LasR activity. This substitution is located in the highly conserved sequence found in many transcription factors, including sigma factors, and may disrupt amphipathic alpha-helix, predicted for the 176 to 189 region, which precedes the putative helix-turn-helix DNA binding motif. We presumed that these three helices may contribute to specific DNA binding.
Fukushima
J
J
Department of Bacteriology, Yokohama City University School of Medicine, Japan.
Ishiwata
T
T
Kurata
M
M
You
Z
Z
Okuda
K
K
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004268:DNA-Binding Proteins / Q000235:genetics*; D010450:Endopeptidases / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial / Q000235:genetics; D014644:Genetic Variation / Q000235:genetics; D018257:Helix-Loop-Helix Motifs; D008969:Molecular Sequence Data; D011550:Pseudomonas aeruginosa / Q000235:genetics*; D017422:Sequence Analysis, DNA; D015534:Trans-Activators / Q000235:genetics*; D015533:Transcriptional Activation
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C080365:LasR protein, Pseudomonas aeruginosa; D015534:Trans-Activators; D010450:Endopeptidases