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Two distinct pathways for cAMP-mediated down-regulation of the beta 2-adrenergic receptor. Phosphorylation of the receptor and regulation of its mRNA level.

Publication ,  Journal Article
Bouvier, M; Collins, S; O'Dowd, BF; Campbell, PT; de Blasi, A; Kobilka, BK; MacGregor, C; Irons, GP; Caron, MG; Lefkowitz, RJ
Published in: J Biol Chem
October 5, 1989

We have studied cyclic AMP-mediated regulation of the beta 2-adrenergic receptor (beta 2AR). The effects of cAMP were assessed in Chinese hamster fibroblast (CHW) cells expressing either the wild type human beta 2AR receptor (CH-beta 2) or mutated forms of the receptor lacking the consensus sequences for phosphorylation by the cAMP-dependent protein kinase. Treatment of the CH-beta 2 cells with the cAMP analogue dibutyryl cAMP (Bt2cAMP) induces a time-dependent "down-regulation" of the number of beta 2AR. This down-regulation of the receptors is accompanied by a decline in the steady state level of beta 2AR mRNA. Moreover, the treatment with Bt2cAMP induces an increase in the phosphorylation level of the membrane-associated beta 2AR. Both the reduction in beta 2AR mRNA and the enhanced phosphorylation of the receptor are rapid and precede the loss of receptor. The down-regulation of beta 2AR induced by Bt2cAMP is concentration-dependent and mimicked by the other biologically active cyclic nucleotide analogue, 8-Br-cAMP, by forskolin, and by the phosphodiesterase inhibitor, isobutylmethylxanthine. In the CHW cell lines expressing receptors lacking the putative protein kinase A phosphorylation sites, the Bt2cAMP-induced phosphorylation of beta 2AR is completely abolished. In these cells the down-regulation of beta 2AR receptor number produced by cAMP is significantly slowed, whereas the reduction in beta 2AR mRNA level is equivalent to that observed in CH-beta 2 cells. These data indicate that there are at least two pathways by which cAMP may decrease the number of beta 2ARs in cells: one involves phosphorylation of the receptor by the cAMP-dependent protein kinase and the other leads to a reduction in steady state beta 2AR mRNA levels.

Duke Scholars

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

October 5, 1989

Volume

264

Issue

28

Start / End Page

16786 / 16792

Location

United States

Related Subject Headings

  • Transfection
  • Transcription, Genetic
  • Theophylline
  • Receptors, Adrenergic, beta
  • RNA, Messenger
  • Phosphorylation
  • Mutation
  • Kinetics
  • Isoproterenol
  • Humans
 

Citation

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Bouvier, M., Collins, S., O’Dowd, B. F., Campbell, P. T., de Blasi, A., Kobilka, B. K., … Lefkowitz, R. J. (1989). Two distinct pathways for cAMP-mediated down-regulation of the beta 2-adrenergic receptor. Phosphorylation of the receptor and regulation of its mRNA level. J Biol Chem, 264(28), 16786–16792.
Bouvier, M., S. Collins, B. F. O’Dowd, P. T. Campbell, A. de Blasi, B. K. Kobilka, C. MacGregor, G. P. Irons, M. G. Caron, and R. J. Lefkowitz. “Two distinct pathways for cAMP-mediated down-regulation of the beta 2-adrenergic receptor. Phosphorylation of the receptor and regulation of its mRNA level.J Biol Chem 264, no. 28 (October 5, 1989): 16786–92.
Bouvier M, Collins S, O’Dowd BF, Campbell PT, de Blasi A, Kobilka BK, et al. Two distinct pathways for cAMP-mediated down-regulation of the beta 2-adrenergic receptor. Phosphorylation of the receptor and regulation of its mRNA level. J Biol Chem. 1989 Oct 5;264(28):16786–92.
Bouvier M, Collins S, O’Dowd BF, Campbell PT, de Blasi A, Kobilka BK, MacGregor C, Irons GP, Caron MG, Lefkowitz RJ. Two distinct pathways for cAMP-mediated down-regulation of the beta 2-adrenergic receptor. Phosphorylation of the receptor and regulation of its mRNA level. J Biol Chem. 1989 Oct 5;264(28):16786–16792.

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

October 5, 1989

Volume

264

Issue

28

Start / End Page

16786 / 16792

Location

United States

Related Subject Headings

  • Transfection
  • Transcription, Genetic
  • Theophylline
  • Receptors, Adrenergic, beta
  • RNA, Messenger
  • Phosphorylation
  • Mutation
  • Kinetics
  • Isoproterenol
  • Humans