Cloning of cDNAs encoding mammalian double-stranded RNA-specific adenosine deaminase.
Double-stranded RNA (dsRNA)-specific adenosine deaminase converts adenosine to inosine in dsRNA. The protein has been purified from calf thymus, and here we describe the cloning of cDNAs encoding both the human and rat proteins as well as a partial bovine clone. The human and rat clones are very similar at the amino acid level except at their N termini and contain three dsRNA binding motifs, a putative nuclear targeting signal, and a possible deaminase motif. Antibodies raised against the protein encoded by the partial bovine clone specifically recognize the calf thymus dsRNA adenosine deaminase. Furthermore, the antibodies can immunodeplete a calf thymus extract of dsRNA adenosine deaminase activity, and the activity can be restored by addition of pure bovine deaminase. Staining of HeLa cells confirms the nuclear localization of the dsRNA-specific adenosine deaminase. In situ hybridization in rat brain slices indicates a widespread distribution of the enzyme in the brain.
O'Connell
M A
MA
Department of Cell Biology, University of Basel, Switzerland.
Krause
S
S
Higuchi
M
M
Hsuan
J J
JJ
Totty
N F
NF
Jenny
A
A
Keller
W
W
D000243:Adenosine Deaminase / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D001921:Brain / Q000201:enzymology; D002417:Cattle; D002467:Cell Nucleus / Q000201:enzymology; D003001:Cloning, Molecular; D003062:Codon; D016384:Consensus Sequence; D018076:DNA, Complementary / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009928:Organ Specificity; D012330:RNA, Double-Stranded / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D016601:RNA-Binding Proteins; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid; D013950:Thymus Gland / Q000201:enzymology*
D003062:Codon; D018076:DNA, Complementary; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; C567955:ADARB1 protein, human; D000243:Adenosine Deaminase