Bovine rotavirus segment 5 protein expressed in the baculovirus system interacts with zinc and RNA

Abstract

The cDNA sequence of genomic segment 5 of bovine rotavirus (RF strain) has been inserted into baculovirus transfer vectors, downstream of the polyhedrin promoter. Recombinant baculoviruses containing gene 5 were selected and the protein was expressed to high yields in Spodoptera frugiperda cells. The recombinant protein was inoculated into rabbits and mice to produce specific hyperimmune antisera. The polyclonal antiserum reacted with a protein in rotavirus-infected MA104 cells and with a protein translated in vitro. This serum was also used to confirm that the gene 5 protein is not a structural protein. Recently, the gene 5 product has been predicted to be a zinc finger protein and reported to contain a highly conserved arrangement of cysteine residues; here, we demonstrate that the recombinant gene 5 protein binds zinc and is an RNA-binding protein as are several other zinc finger proteins.