Purification and characterization of a human RNA adenosine deaminase for glutamate receptor B pre-mRNA editing.
The glutamate receptor subunit B (GluR-B) pre-mRNA is edited at two adenosine residues, resulting in amino acid changes that alter the electrophysiologic properties of the glutamate receptor. Previous studies showed that these amino acid changes are due to adenosine to inosine conversions in two codons resulting from adenosine deamination. Here, we describe the purification and characterization of an activity from human HeLa cells that efficiently and accurately edits GluR-B pre-mRNA at both of these sites. The purified activity contains a human homolog of the recently reported rat RED1 (rRED1) protein, a member of the family of double-stranded RNA-dependent deaminase proteins. Recombinant human RED1 (hRED1), but not recombinant dsRAD, another member of the family, efficiently edits both the Q/R and R/G sites of GluR-B RNA. We conclude that the GluR-B editing activity present in HeLa cell extracts and the recombinant hRED1 protein are indistinguishable.
Yang
J H
JH
Department of Molecular and Cellular Biology, Harvard University, 7 Divinity Avenue, Cambridge, MA 02138, USA.
Sklar
P
P
Axel
R
R
Maniatis
T
T
D000241:Adenosine / Q000378:metabolism; D000243:Adenosine Deaminase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D006367:HeLa Cells; D006801:Humans; D007288:Inosine / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D017393:RNA Editing; D012322:RNA Precursors / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins; D017470:Receptors, Glutamate / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity
D012322:RNA Precursors; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D017470:Receptors, Glutamate; D011994:Recombinant Proteins; D007288:Inosine; C567955:ADARB1 protein, human; D000243:Adenosine Deaminase; D000241:Adenosine