VP39 is a bifunctional vaccinia virus protein that acts as both an mRNA cap–specific RNA 2′-O-methyltransferase and a poly(A) polymerase processivity factor. Here, we report the 1.85 Å crystal structure of a VP39 variant complexed with its AdoMet cofactor. VP39 comprises a single core domain with structural similarity to the catalytic domains of other methyltransferases. Surface features and mutagenesis data suggest two possible RNA-binding sites with novel underlying architecture, one of which forms a cleft spanning the region adjacent to the methyltransferase active site. This report provides a prototypic structure for an RNA methyltransferase, a protein that interacts with the mRNA 5′ cap, and an intact poxvirus protein.
