Mutational analysis of the Chlamydia trachomatis dnaK promoter defines the optimal –35 promoter element

A long-standing question in the biology of the intracellular bacterium, Chlamydia, has been the structure of the promoter recognized by its RNA polymerase. The ‘RNA polymerase sigma subunit paradox’ refers to the difficulty reconciling the conservation between the RNA polymerases of Chlamydia and Escherichia coli, especially at the level of the promoter-recognition sigma subunit, with the general lack of homology between chlamydial promoters and the E.coli σ(70) consensus promoter. While the –10 promoter element appears to be conserved between Chlamydia and E.coli, the structure of the chlamydial –35 promoter element has not been defined. We have investigated the structure of the –35 element of the Chlamydia trachomatis dnaK promoter by measuring the effects of single base pair substitutions on in vitro promoter activity. Most substitutions produced large decreases in promoter activity, which allowed us to define the optimal –35 sequence in the context of the dnaK promoter. We found that the optimal chlamydial –35 promoter sequence is identical to the E.coli σ(70) consensus –35 promoter element (TTGACA). These results indicate that the optimal promoter specificities of the major form of chlamydial RNA polymerase and E.coli σ(70) RNA polymerase are in fact highly conserved. A further implication of our results is that many chlamydial promoters have a suboptimal promoter structure. We hypothesize that these chlamydial promoters are intrinsically weak promoters that can be regulated during the chlamydial developmental cycle by additional transcription factors.