Acquisition of apparent DNA slippage structures during extensive evolutionary divergence of pcaD and catD genes encoding identical catalytic activities in Acinetobacter calcoaceticus.

The pca operon from the Gram- bacterium Acinetobacter calcoaceticus encodes all of the enzymes required for catabolism of protocatechuate to common intermediary metabolites. This report presents the 2754-nucleotide (nt) sequence of a HindIII restriction fragment containing pcaD, the 801-bp gene encoding beta-ketoadipate enol-lactone hydrolase I. The deduced primary structure of A. calcoaceticus PcaD shares 44% amino acid (aa) sequence identity with the aligned primary structure of CatD (beta-ketoadipate enol-lactone hydrolase II) from the same organism, and the overall nt sequence identity of the two genes is 51.8%. In the 56% of the genes where selection for identical aa residues was not imposed, pcaD and catD have diverged so extensively that nt sequence identity of the aligned segments is only 28.2%; the G+C contents of these segments from the respective genes differ by 8%. Conserved within the aligned PcaD and CatD aa sequences is a Ser residue corresponding to the nucleophile within the alpha/beta-fold of many hydrolytic enzymes. In this region of primary structure, PcaD and CatD appear to have maintained some different aa sequences derived from a common ancestor. Conservation of the different aa sequences during extreme evolutionary divergence suggests that separate segments of primary structure, conserved within either PcaD or CatD, may be functionally incompatible within recombinant enzymes. Consequently, selection for avoidance of genetic exchange between pcaD and catD could account for the thorough nt substitution in regions where identical aa were not selected. Sequence repetitions within pcaD suggest that the multiple mutations required for its extensive divergence from catD were achieved in part by acquisition of a complex DNA slippage structure.