Expression of mRNA of β1- and β2-adrenergic receptors in Xenopus oocytes results from structurally distinct receptor mRNAs

Suleiman Bahouth, J. R. Hadcock, C. C. Malbon

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Poly(A)+-selected RNA prepared from cells or tissues that express a homogeneous population of either β1- or β2-adrenergic receptors was isolated and then microinjected into Xenopus laevis oocytes. Following microinjection, the expression of β-adrenergic receptors was assessed by equilibrium radioligand binding analysis using the antagonist ligand [3H]dihydroalprenolol. The pharmacology of theh newly- expressed β-adrenergic receptors in oocyte membranes was the same as that of the original tissue used as a source of RNA. Hybridization of nick-translated cDNA of hamster β2-adrenergic receptor to poly(A)+-selected RNA from tissues containing β2-adrenergic receptors was to a mRNA species of 2.2 kilobases. In contrast, hybridization of the cDNA probe to poly(A)+-selected RNA from tissues containing β1-adrenergic receptors was to a mRNA species of 2.0 kilobases. A single-stranded fragment of hamster β2-adrenergic receptor cDNA corresponding to nucleotides 730-886 was isolated and uniformly radiolabeled. This region of the gene is predicted to encode for the entire second exofacial loop (L4-5), the entire fifth transmembrane-spanning region, and the first 5 amino acid residues of the third cytoplasmic loop (L5-6) of the β2-adrenergic receptor. Hybridization at 48 and 56 °C of poly(A)+-selected RNA prepared from sources that express either β1-or β2-adrenergic receptors to the antisense orientation strand of this region of the β2-adrenergic receptor cDNA was followed by S1 endonuclease digestion of nonhybridized sequences. At 48 °C, S1-resistant hybrids from both sources of RNA protected the probe from S1 endonuclease digestion. At 56 °C, however, only the RNA prepared from the source of β2-adrenergic receptors protected the probe from S1 endonuclease digestion. These results demonstrate that the mRNAs encoding for the structurally homologous β1- and β2-adrenergic receptors are distinct in the pharmacological specificity of their translation products and in their size and structure.

Original languageEnglish (US)
Pages (from-to)8822-8826
Number of pages5
JournalJournal of Biological Chemistry
Volume263
Issue number18
StatePublished - 1988
Externally publishedYes

Fingerprint

Xenopus
Adrenergic Receptors
Oocytes
Messenger RNA
Endonucleases
Complementary DNA
Tissue
Digestion
RNA
Cricetinae
Dihydroalprenolol
Pharmacology
RNA Probes
Xenopus laevis
Microinjections
Nucleotides
Genes
Ligands
Membranes
Amino Acids

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Expression of mRNA of β1- and β2-adrenergic receptors in Xenopus oocytes results from structurally distinct receptor mRNAs. / Bahouth, Suleiman; Hadcock, J. R.; Malbon, C. C.

In: Journal of Biological Chemistry, Vol. 263, No. 18, 1988, p. 8822-8826.

Research output: Contribution to journalArticle

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