IDENTITY OF THE RNA-BINDING PROTEIN-K OF HNRNP PARTICLES WITH PROTEIN-H16, A SEQUENCE-SPECIFIC SINGLE-STRAND DNA-BINDING PROTEIN

Protein H16, which we have identified previously in mammalian cell lin es, binds in vitro to two single stranded DNA sites on the late strand of the early promoter of SV40. It has no other single strand binding site in the SV40 genome and does not bind to double stranded DNA. In v itro, H16 can be shown to stimulate strongly the activity of purified RNA polymerase II. Here we have purified this 70 kDa protein from cult ured monkey cells and have sequenced three of its tryptic peptides. Th e analysis indicates that H16 is the simian homolog of human protein K , a nuclear RNA-binding protein found in heterogeneous nuclear ribonuc leo-protein (hnRNP) particles, which contains a KH domain present in s everal proteins including the fragile X mental retardation gene produc t (FMR1). The binding affinities of protein K/H16 for RNA and DNA were subsequently compared in detail. They showed that under conditions wh ere K/H16 binds strongly to its single stranded DNA site, it binds ver y weakly to the corresponding RNA sequence. This result suggests a pos sible shuttling of the protein from RNA to DNA during processes which involve opening of the DNA double helix.