Supplementary material from "Identification of a novel nuclease activity in human DDX49 helicase"

Human DDX49 is an emerging target to understand cancer progression and retroviral diseases through its essential roles in nucleolar RNA processing. Here, we identify nuclease activity of human DDX49, which requires a Asp-X-Asp active site that is conserved across the metazoan DDX49s, but is absent from DDX49 homologues in yeast and archaea. We provide evidence that DDX49 RNA helicase activity facilitates the nuclease activity. Using CRISPR-Cas9 genetic editing we show that a heterozygous (DDX49+/-)U2OS cell line is defective at cell migration, a phenotype supporting the association of DDX49 with cancer cell invasiveness. Measurement of RNAs in DDX49+/- indicate that DDX49 is required to sustain levels of 5.8S rRNA.