Molecular evolution and phylogenetic utility of the polyubiquitin locus in mammals and higher vertebrates.

The product of ubiquitin genes is a small protein involved in intracellular sorting of other proteins. The locus consists of tandemly arrayed, uninterrupted copies of the gene. As several studies have noted, the Polyubiquitin locus is a model system for studying concerted evolution. While the protein is among the most conserved known, individual copies within an organism show variation in nucleotide sequence despite clear evidence of concerted evolution. When treated as individuals, repeats from a given locus form a monophyletic group. Furthermore adjacent copies often cluster, suggestive of the mechanism of concerted evolution. Due to this concerted evolution of repeats (and loci in organisms with multiple polyubiquitins), sequencing of heterogeneous PCR products consisting of all the repeats in a given organism may yield phylogenetic signal, as with other multicopy genes. We test this possibility through 22 original sequences using primers designed so that only tandem copies are amplified. Using these and previously published data, we further explore these phenomena in higher vertebrates and mammals in particular. We suggest that multiple locus duplications have occurred within mammals. Positional codon bias is strongly evident. We also find substitutional bias with regard to codon type. GC content of the locus appears to be generally high across vertebrates. Intraorganismal variation is tallied as an indication of frequency of change in codon position and transition/transversion ratios to further elucidate the tempo and mode of molecular evolution. Using these data, a weighting scheme for ubiquitin is also presented. Despite the gene's high GC content, transitional changes still appear more frequent. While the phylogenetic utility of ubiquitin does not appear great, its mechanistic insights seem far from exhausted.