Patterns of nucleotide difference in overlapping and non-overlapping reading frames of papillomavirus genomes.

On the basis of a phylogenetic analysis of papillomavirus complete genomes, 30 pairs of closely related genomes were chosen for examination of the pattern of synonymous and non-synonymous nucleotide difference in overlapping and non-overlapping reading frames. The proportion of synonymous differences per synonymous site (pS) was generally reduced in overlapping regions in comparison to non-overlapping regions. This reduction in pS appears to result from purifying selection acting on the overlapping reading frame because of functional constraint on the amino acid sequence that it encodes. As a consequence of such purifying selection, a trade-off was observed between synonymous substitution in one reading frame and non-synonymous substitution in the other; this pattern was particularly evident in the overlap between the E2 and E1 reading frames, which encodes a highly conserved portion of the E2 protein. Fifty-three sequences of the hinge region of the HPV16 E2 protein (where the E2 and E4 reading frames overlap) showed an elevated level of non-synonymous nucleotide substitution in the E2 reading frame, particularly in a region believed to be involved in immune recognition. Non-synonymous substitutions in this region occurred disproportionately in such a way as to cause synonymous substitutions in the E4 reading frame. Thus, positive diversifying selection on the E2 hinge region appears to coexist with purifying selection on the overlapping region of E4.