Isolation and characterization of an avian A1 adenosine receptor gene and a related cDNA clone.
We have isolated the gene for a chick A1 adenosine receptor along with a cDNA that codes for the same adenosine receptor. The cDNA clone was isolated from both adipose tissue and heart cDNA libraries and encodes a 324-amino acid protein with 80% identity with mammalian A1 adenosine receptors. Transient expression of the cDNA in human embryonic kidney (HEK) 293 cells shows that it encodes a protein that binds [3H]CCPA (2-chloro-N6-[cyclopentyl-2,3,4,5-3H]cyclopentyladenosine, a specific agonist radioligand, with a KD of 5.6 +/- 2.4 nM. Cyclic AMP measurements in HEK 293 cells co-transfected with the chick cDNA and a cDNA for a luteinizing hormone/choriogonadotropin receptor shows that A1 adenosine receptor agonists antagonize the cyclic AMP-elevating effect of bovine luteinizing hormone. Two partial genomic clones were isolated. The first contains 5'-untranslated sequence including a putative promoter region which does not contain a TATA box, an intron and the first third of the coding sequence of the A1 adenosine receptor cDNA. The coding sequence of this partial genomic clone terminates at a second intron. The second partial genomic clone contains the rest of the coding sequence and the 3'-untranslated elements in a single exon. Thus the chick A1 adenosine receptor gene contains one intron in the 5'-untranslated region and a minimum of one intron in the coding sequence.
Aguilar
J S
JS
Department of Pharmacology, School of Medicine, University of Illinois at Chicago, USA.
Tan
F
F
Durand
I
I
Green
R D
RD
D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001923:Brain Chemistry; D002645:Chickens; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000378:metabolism; D015342:DNA Probes; D018076:DNA, Complementary / Q000235:genetics* / Q000302:isolation & purification*; D004285:Dogs; D005784:Gene Amplification; D015870:Gene Expression; D016678:Genome; D007438:Introns; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D018047:Receptors, Purinergic P1 / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic; D014162:Transfection
D015342:DNA Probes; D018076:DNA, Complementary; D018047:Receptors, Purinergic P1; C058394:2-chloro-N(6)cyclopentyladenosine; D000242:Cyclic AMP; D000241:Adenosine