Comparison of relative fixation rates of synonymous (silent) and nonsynonym
ous (amino acid-altering) mutations provides a means for understanding the
mechanisms of molecular sequence evolution. The nonsynonymous/synonymous ra
te ratio (omega = d(N)/d(S)) is an important indicator of selective pressur
e at the protein level, with omega = 1 meaning neutral mutations, omega < 1
purifying selection, and omega > 1 diversifying positive selection. Amino
acid sites in a protein are expected to be under different selective pressu
res and have different underlying omega ratios. We develop models that acco
unt for heterogeneous omega ratios among amino acid sites and apply them to
phylogenetic analyses of protein-coding DNA sequences. These models are us
eful for testing for adaptive molecular evolution and identifying amino aci
d sites under diversifying selection. Ten data sets of genes from nuclear,
mitochondrial, and viral genomes are analyzed to estimate the distributions
of omega among sites. In all data sets analyzed, the selective pressure in
dicated by the omega ratio is found to be highly heterogeneous among sites.
Previously unsuspected Darwinian selection is detected in several genes in
which the average omega ratio across sites is <1, but in which some sites
are clearly under diversifying selection with omega > 1. Genes undergoing p
ositive selection include the beta-globin gene from vertebrates, mitochondr
ial protein-coding genes from hominoids, the hemagglutinin (HA) gene from h
uman influenza virus A, and HIV-1 env, vif, and pol genes. Tests for the pr
esence of positively selected sites and their subsequent identification app
ear quite robust to the specific distributional form assumed for omega and
call be;achieved using any of several models we implement. However, we enco
untered difficulties in estimating the precise distribution of omega among
sites from real data sets.