Adaptive diversification within a large family of recently duplicated, placentally expressed genes

The pregnancy-associated glycoproteins (PAC) are putative peptide-binding p roteins and products of a large family of genes whose expression is localiz ed to the placental surface epithelium of artiodactyl species. We have test ed the hypothesis that natural selection has favored diversification of the se genes by examining patterns of nucleotide substitution in a sample of 28 closely related bovine. caprine, and ovine family members that are express ed only in trophoblast binucleate cells. Three observations were made. Firs t, in codons encoding highly variable domains of the proteins, there was a greater accumulation of both synonymous and nonsynonymous mutations than in the more conserved regions of the genes. Second, in the variable regions. the mean number of nonsynonymous nucleotide substitutions per site was sign ificantly greater than the mean number of synonymous substitutions per site . Third, nonsynonymous changes affecting amino acid charge occurred more fr equently than expected under random substitution. This unusual pattern of n ucleotide substitution implies that natural selection has acted to diversif y these PAC molecules at the amino acid level, which in turn suggests that these molecules have undergone functional diversification. We estimate that the binucleate cell-expressed PAC originated 52 +/- 6 million years ago, s oon after the divergence of the ruminant lineage. Thus, rapid functional di versification of PAG expressed in trophoblast binucleate cells seems to hav e been associated with the origin of this unique placental adaptation.