DRBP76, a double-stranded RNA-binding nuclear protein, is phosphorylated by the interferon-induced protein kinase, PKR.
The interferon-induced double-stranded RNA-activated protein kinase PKR is the prototype of a class of double-stranded (dsRNA)-binding proteins (DRBPs) which share a dsRNA-binding motif conserved from Drosophila to humans. Here we report the purification of DRBP76, a new human member of this class of proteins. Sequence from the amino terminus of ... DRBP76 matched that of the M phase-specific protein, MPP4. DRBP76 was also cloned by the yeast two-hybrid screening of a cDNA library using a mutant PKR as bait. Analysis of the cDNA sequence revealed that it is the full-length version of MPP4, has a bipartite nuclear localization signal, two motifs that can mediate interactions with both dsRNA and PKR, five epitopes for potential M phase-specific phosphorylation, two potential sites for phosphorylation by cyclin-dependent kinases, a RG2 motif present in many RNA-binding proteins and predicts a protein of 76 kDa. DsRNA and PKR interactions of DRBP76 were confirmed by analysis of in vitro translated and purified native proteins. Cellular expression of an epitope-tagged DRBP76 demonstrated its nuclear localization, and its co-immunoprecipitation with PKR demonstrated that the two proteins interact in vivo. Finally, purified DRBP76 was shown to be a substrate of PKR in vitro, indicating that this protein's cellular activities may be regulated by PKR-mediated phosphorylation.
Mesh Terms:
Amino Acid Sequence, Base Sequence, Cell Cycle, DNA, Complementary, Enzyme Induction, Hela Cells, Humans, Interferons, Molecular Sequence Data, Nuclear Factor 90 Proteins, Nuclear Proteins, Phosphoproteins, Phosphorylation, Protein Binding, RNA, Double-Stranded, RNA-Binding Proteins, eIF-2 Kinase
Amino Acid Sequence, Base Sequence, Cell Cycle, DNA, Complementary, Enzyme Induction, Hela Cells, Humans, Interferons, Molecular Sequence Data, Nuclear Factor 90 Proteins, Nuclear Proteins, Phosphoproteins, Phosphorylation, Protein Binding, RNA, Double-Stranded, RNA-Binding Proteins, eIF-2 Kinase
J. Biol. Chem.
Date: Jul. 16, 1999
PubMed ID: 10400669
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