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Ingrid Chou Koo

Publications and source records attributed to Ingrid Chou Koo.

2 recordsLinked to original sources

ChxR is a transcriptional activator in Chlamydia.

Chlamydia spp. are obligate intracellular bacterial pathogens that alternate between two metabolically and morphologically distinct developmental forms, and differentiation depends on transcriptional regulation. Genome sequencing of Chlamydia trachomatis revealed an ORF, CT630 (chxR), whose amino acid sequence contains a winged helix-turn-helix motif similar to the DNA-binding domain of response regulators in the OmpR subfamily. ChxR differs from many response regulators in that essential residues in the receiver or phosphorylation domain are lacking. ChxR functions as a transcriptional regulator because it activated transcription of ompF and ompC when expressed in Escherichia coli. In vitro transcription combined with microarray analysis also demonstrated that ChxR activates its own expression by binding directly to sites upstream of chxR; it also activates infA, tufA, oppA, and CT084. DNase I protection studies showed that ChxR bound to sites in the ompF and ompC promoter proximal regions that overlap but were distinct from OmpR binding sites. Both proteins could bind simultaneously to their nonoverlapping binding sites. This report identifies a stage-specific transcriptional regulator and some of its target genes in Chlamydia.

Bacterial Proteins↗

A developmentally regulated two-component signal transduction system in Chlamydia.

Two-component systems allow bacteria to adapt to changing environmental conditions and may induce developmental changes necessary for survival. Chlamydia trachomatis alternates between two distinct developmental forms, each optimized for survival in a separate niche. Transcriptional regulation of development is not understood. The C. trachomatis genome sequence revealed a single pair of genes (ctcB-ctcC) predicted to encode proteins with sequence conservation to bacterial two-component systems. Sequence analysis revealed that the sensor kinase, CtcB, possessed an energy-sensing PAS domain and phosphorylation site. The response regulator, CtcC, had homology to sigma(54) activators, possessing conserved receiver and ATPase domains and phosphorylation site, but lacked the C-terminal DNA-binding domain. ctcB and ctcC were expressed late in the developmental cycle, and both proteins were detected in EB lysates. Recombinant CtcB and CtcC were purified from denatured Escherichia coli inclusion bodies and refolded. CtcC was found to aggregate as dimers and tetramers in solution. In vitro phosphorylation assays showed that CtcB autophosphorylated in the presence of Mg(2+), Mn(2+), and Fe(2+) and transferred the phosphoryl group in the presence of CtcC. Collectively, these results show that CtcB and CtcC function as a two-component system and are likely responsible for transcriptional regulation by sigma(54) holoenzyme during late-stage chlamydial development.

Amino Acid Sequence↗