Substrate specificity of the dsRNA unwinding/modifying activity.
Double-stranded RNA (dsRNA) unwinding/modifying activity, which is present in a wide range of eukaryotic cells, has been previously shown to convert up to 50% of adenosine residues to inosines within intermolecular dsRNA. In the present study, we report that this activity also modifies, though slightly less efficiently, intramolecular double-stranded regions of synthetic RNAs. Our results widen the range of the possible biological substrates for the activity since many stem and loop type RNA secondary structures (intramolecular dsRNA), present in eukaryotic as well as viral transcripts, can potentially serve as substrates. In addition, we have found that the dsRNA unwinding/modifying activity requires a double-stranded region of at least 15-20 base pairs (bp) for substrate recognition. Furthermore, modification efficiency was found to be critically dependent on the length of the double-stranded region; as the size decreased below 100 bp, it dropped precipitously. Our results suggest that efficient modification may occur only with relatively long (greater than 100 bp) dsRNA, perhaps because multiple copies of the enzyme must be bound.
Wistar Institute of Anatomy and Biology, Philadelphia, PA 19104.
K
K
Nishikura
C
C
Yoo
U
U
Kim
J M
JM
Murray
P A
PA
Estes
F E
FE
Cash
S A
SA
Liebhaber
D001483:Base Sequence; D001667:Binding, Competitive; D004247:DNA / Q000235:genetics; D005914:Globins / Q000235:genetics; D013006:Growth Hormone / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010957:Plasmids; D012330:RNA, Double-Stranded / Q000235:genetics / Q000378:metabolism*
D012330:RNA, Double-Stranded; D013006:Growth Hormone; D005914:Globins; D004247:DNA