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Control of Gene Expression by Catecholamines and the Renin-Angiotensin System pp 73–79Cite as

Catecholamines and angiotensinogen gene expression in kidney proximal tubular cells

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Part of the book series: Developments in Molecular and Cellular Biochemistry ((DMCB,volume 33))

Abstract

To investigate the molecular mechanism(s) of action of catecholamines on the expression of the angiotensinogen (ANG) gene in kidney proximal tubular cells, we used opossum kidney (OK) cells with a fusion gene containing the 5’-flanking regulatory sequence of the rat ANG gene fused with a human growth hormone (hGH) gene as a reporter, pOGH (rANG N-1498/+18), permanently integrated into their genomes. The level of expression of the ANG-GH fusion gene was quantified by the amount of immunoreactive-hGH (IR-hGH) secreted into the medium. The addition of norepinephrine (NE), isoproterenol (a β1/β2-adrenergic receptor (AR) agonist) and iodoclonidine (an α2-AR agonist) stimulated the expression of the ANG-GH fusion gene in a dose-dependent manner, whereas the addition of epinephrine and phenylephrine (α1-AR agonist) had no effect. The stimulatory effect of NE was blocked by the presence of propranolol (β-AR blocker), atenolol (β1-AR blocker), yohimbine (α2-AR blocker), Rp-cAMP (an inhibitor of cAMP-dependent protein kinase AI & AII) and staurosporine (an inhibitor of protein kinase C), but was not blocked by ICI 118, 551 (β2-AR blocker) and prazosin (α1-AR blocker). The addition of a combination of isoproterenol and iodoclonidine or a combination of 8-Bromo-cAMP (8-Br-cAMP) and phorbol 12-myristate (PMA) synergistically stimulated the expression of the ANG-GH fusion gene as compared to the addition of isoproterenol, iodoclonidine, 8-Br-cAMP or PMA alone. Furthermore, the addition of NE, 8-Br-cAMP or PMA stimulated the expression of pOGH (rANG N-806/-779/ -53/+18), a fusion gene containing the putative cAMP responsive element (CRE, ANG N-806/-779) upstream of the ANG promoter (ANG N-53/+1 8) in OK cells, but had no effect on the expression of fusion genes containing the mutant of the CRE. Gel mobility shift assays revealed that the ANG-CRE binds with the DNA-binding domain (bZIP 254-327) of the cAMP-responsive binding protein (CREB). The binding of the labeled ANG-CRE to CREB (bZIP254-327) was displaced by unlabeled ANGCRE and the CRE of the somatostatin gene but not by the mutants of the ANG-CRE. Finally, NE stimulated the phosphorylation of CREB in OK cells. These studies demonstrate that the molecular mechanism(s) of NE action on the expression of the ANG gene in OK cells may be mediated via both the PKA and PKC signalling pathways and via the phosphorylation of CREB. The phosphorylated CREB then interacts with the CRE in the 5’-flanking region of the ANG gene and subsequently stimulates the gene expression. (Mol Cell Biochem 212: 73–79, 2000)

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Abbreviations

AC:

adenyllyl cyclase

ANG:

angiotensinogen

Ang I:

angiotensin I

Ang II:

angiotensin II

ACE:

angiotensin converting enzyme

AR:

adrenergic receptor

CRE:

cyclic AMP responsive element

CREB:

cyclic AMP responsive element binding protein

NE:

norepinephrine

OK:

opssum kidney

PKA:

protein kinase A

PKC:

protein kinase C

RAS:

renin-angiotensin system

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Authors and Affiliations

  1. Maisonneuve-Rosemont Hospital Research Center, University of Montreal, Montreal, Quebec, Canada

    John S. D. Chan, Tian-Tian Wang, Shao-Ling Zhang & Xing Chen

  2. Servier Amérique, Laval, Quebec, Canada

    Serge Carrière

  3. Maisonneuve-Rosemont Hospital Research Center, University of Montreal, 5415 Boulevard de l’Assomption, Montreal, Quebec, Canada, H1T 2M4

    John S. D. Chan

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  1. John S. D. Chan
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  4. Xing Chen
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  5. Serge Carrière
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  1. Internal Medicine and Cardiology, Molecular Cardiology Laboratory, Philipps University of Marburg, Karl-von-Frisch-Strasse 1, 35044, Marburg, Germany

    Heinz Rupp

  2. Internal Medical, Molecular Cardiology Laboratory, Philipps University of Marburg, Karl-von-Frisch-Strasse, 35033, Marburg, Germany

    Bernard Maisch

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Chan, J.S.D., Wang, TT., Zhang, SL., Chen, X., Carrière, S. (2000). Catecholamines and angiotensinogen gene expression in kidney proximal tubular cells. In: Rupp, H., Maisch, B. (eds) Control of Gene Expression by Catecholamines and the Renin-Angiotensin System. Developments in Molecular and Cellular Biochemistry, vol 33. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-4351-0_9

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