Molecular basis defining human Chlamydia trachomatis tissue tropism. A possible role for tryptophan synthase

• Published in: The Journal of biological chemistry Vol. 277; no. 30; pp. 26893 - 26903
• Main Authors: Fehlner-Gardiner, Christine, Roshick, Christine, Carlson, John H, Hughes, Scott, Belland, Robert J, Caldwell, Harlan D, McClarty, Grant
• Format: Journal Article
• Language: English
• Published: United States 26.07.2002
• Subjects: Amino Acid Sequence; Binding Sites; Blotting, Western; Cells, Cultured; Chlamydia trachomatis - enzymology; Chlamydia trachomatis - physiology; Cloning, Molecular; Escherichia coli - metabolism; Genetic Complementation Test; Glycerophosphates - metabolism; HeLa Cells; Humans; Models, Biological; Molecular Sequence Data; Mutation; Plasmids - metabolism; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - metabolism; Salmonella - metabolism; Sequence Homology, Amino Acid; Transcription, Genetic; Tryptophan - metabolism; Tryptophan Synthase - physiology
• ISSN: 0021-9258
• DOI: 10.1074/jbc.M203937200

Here we report the cloning and sequencing of a region of the chlamydiae chromosome termed the "plasticity zone" from all the human serovars of C. trachomatis containing the tryptophan biosynthesis genes. Our results show that this region contains orthologues of the tryptophan repressor as well as the alpha and beta subunits of tryptophan synthase. Results from reverse transcription-PCR and Western blot analyses indicate that the trpBA genes are transcribed, and protein products are expressed. The TrpB sequences from all serovars are highly conserved. In comparison with other tryptophan synthase beta subunits, the chlamydial TrpB subunit retains all conserved amino acid residues required for beta reaction activity. In contrast, the chlamydial TrpA sequences display numerous mutations, which distinguish them from TrpA sequences of all other prokaryotes. All ocular serovars contain a deletion mutation resulting in a truncated TrpA protein, which lacks alpha reaction activity. The TrpA protein from the genital serovars retains conserved amino acids required for catalysis but has mutated several active site residues involved in substrate binding. Complementation analysis in Escherichia coli strains, with defined mutations in tryptophan biosynthesis, and in vitro enzyme activity data, with cloned TrpB and TrpA proteins, indicate these mutations result in a TrpA protein that is unable to utilize indole glycerol 3-phosphate as substrate. In contrast, the chlamydial TrpB protein can carry out the beta reaction, which catalyzes the formation of tryptophan from indole and serine. The activity of the chlamydial Trp B protein differs from that of the well characterized E. coli and Salmonella TrpBs in displaying an absolute requirement for full-length TrpA. Taken together our data indicate that genital, but not ocular, serovars are capable of utilizing exogenous indole for the biosynthesis of tryptophan.