Origins and Molecular Features of a Chimeric Type-P4A ATPase-Guanylate Cyclase from Stramenopiles and Alveolates

A chimeric protein produced from the fusion of a type-P4 ATPase, also called flippase, and a guanylate cyclase (P4GC) was originally described in ciliates and malaria parasites. An extensive search for P4GC homologs was carried out, and the protein's structural features were determined by sequence alignments and homology modeling. P4GC is found in all alveolate lineages and in some stramenopile lineages. This suggests that the gene fusion event occurred in a common ancestor of stramenopiles and alveolates after the divergence of rhizarians, although alternative evolutionary scenarios cannot be excluded. This scenario necessitates the subsequent loss of P4GC in some stramenopile lineages. Alternatively, the gene fusion may have arisen in an early alveolate lineage and subsequently been transferred horizontally to an ancestor of oomycetes and bicosoecids, or vice versa. The sequence homology of the flippase module is not well preserved in some lineages, whereas the structure and sequence homology of the cyclase module is highly conserved in all lineages. These findings suggest that the cyclase module has been subject to stronger evolutionary constraints than the flippase module, while the latter may have undergone lineage-specific functional diversification. However, despite sequence divergence, homology modeling indicates that the predicted 3-dimensional structures of the flippase modules are highly conserved across all lineages, suggesting that structural constraints have preserved its functional role(s) within P4GC. Together, these findings indicate that P4GC has undergone lineage-specific functional diversification while maintaining a highly conserved guanylate cyclase module throughout alveolate and stramenopile evolution.