Viral proteins originated de novo by overprinting can be identified by codon usage: application to the 'gene nursery' of Deltaretroviruses

A well-known mechanism through which new protein-coding genes originate is by modification of pre-existing genes, e.g. by duplication or horizontal transfer. In contrast, many viruses generate protein-coding genes de novo, via the overprinting of a new reading frame onto an existing (“ancestral”) frame. This mechanism is thought to play an important role in viral pathogenicity, but has been poorly explored, perhaps because identifying the de novo frames is very challenging. Therefore, a new approach to detect them was needed. We assembled a reference set of overlapping genes for which we could reliably determine the ancestral frames, and found that their codon usage was significantly closer to that of the rest of the viral genome than the codon usage of de novo frames. Based on this observation, we designed a method that allowed the identification of de novo frames based on their codon usage with a very good specificity, but intermediate sensitivity. Using our method, we predicted that the Rex gene of deltaretroviruses has originated de novo by overprinting the Tax gene. Intriguingly, several genes in the same genomic region have also originated de novo and encode proteins that regulate the functions of Tax. Such “gene nurseries” may be common in viral genomes. Finally, our results confirm that the genomic GC content is not the only determinant of codon usage in viruses and suggest that a constraint linked to translation must influence codon usage.
Author Summary How does novelty originate in nature? It is commonly thought that new genes are generated mainly by modifications of existing genes (the “tinkering” model). In contrast, we have shown recently that in viruses, numerous genes are generated entirely de novo (“from scratch”). The role of these genes remains underexplored, however, because they are difficult to identify. We have therefore developed a new method to detect genes originated de novo in viral genomes, based on the observation that each viral genome has a unique “signature”, which genes originated de novo do not share. We applied this method to analyze the genes of Human T-Lymphotropic Virus 1 (HTLV1), a relative of the HIV virus and also a major human pathogen that infects about twenty million people worldwide. The life cycle of HTLV1 is finely regulated – it can stay dormant for long periods and can provoke blood cancers (leukemias) after a very long incubation. We discovered that several of the genes of HTLV1 have originated de novo. These novel genes play a key role in regulating the life cycle of HTLV1, and presumably its pathogenicity. Our investigations suggest that such “gene nurseries” may be common in viruses.