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. 1994 Aug 15;302(Pt 1):81–86. doi: 10.1042/bj3020081

Stabilization of the mRNA for the uncoupling protein thermogenin by transcriptional/translational blockade and by noradrenaline in brown adipocytes differentiated in culture: a degradation factor induced by cessation of stimulation?

C Picó 1, D Herron 1, A Palou 1, A Jacobsson 1, B Cannon 1, J Nedergaard 1
PMCID: PMC1137193  PMID: 8068027

Abstract

The stability of the mRNA coding for the uncoupling protein thermogenin was investigated in mouse brown-fat cells differentiated in culture. After 7 days in culture, the cells were stimulated for 24 h with noradrenaline, and a high level of thermogenin mRNA was then observed. If noradrenaline treatment was continued, the mRNA level remained high, but, upon withdrawal of noradrenaline, the level decreased rapidly, with a half-life of only 2.7 h. The presence of transcriptional (actinomycin) or translational (cycloheximide) inhibitors prolonged the apparent half-life by about 50%. The presence of noradrenaline during transcriptional blockade led to a further stabilization of thermogenin mRNA. It was concluded that an induced (or short-lived) gene product is important for thermogenin mRNA degradation. Direct interaction of noradrenaline with the cultured brown adipocytes could apparently not mimic the paradoxical destabilization of thermogenin mRNA in vivo, previously observed in the cold-exposed mouse [Jacobsson, Cannon and Nedergaard (1987) FEBS Lett. 244, 353-356], indicating significant differences between the systems in vitro and in vivo.

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Selected References

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  1. Atwater J. A., Wisdom R., Verma I. M. Regulated mRNA stability. Annu Rev Genet. 1990;24:519–541. doi: 10.1146/annurev.ge.24.120190.002511. [DOI] [PubMed] [Google Scholar]
  2. Bernstein P., Ross J. Poly(A), poly(A) binding protein and the regulation of mRNA stability. Trends Biochem Sci. 1989 Sep;14(9):373–377. doi: 10.1016/0968-0004(89)90011-x. [DOI] [PubMed] [Google Scholar]
  3. Bouillaud F., Ricquier D., Mory G., Thibault J. Increased level of mRNA for the uncoupling protein in brown adipose tissue of rats during thermogenesis induced by cold exposure or norepinephrine infusion. J Biol Chem. 1984 Sep 25;259(18):11583–11586. [PubMed] [Google Scholar]
  4. Bouillaud F., Ricquier D., Thibault J., Weissenbach J. Molecular approach to thermogenesis in brown adipose tissue: cDNA cloning of the mitochondrial uncoupling protein. Proc Natl Acad Sci U S A. 1985 Jan;82(2):445–448. doi: 10.1073/pnas.82.2.445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brawerman G. mRNA decay: finding the right targets. Cell. 1989 Apr 7;57(1):9–10. doi: 10.1016/0092-8674(89)90166-9. [DOI] [PubMed] [Google Scholar]
  6. Bronnikov G., Houstek J., Nedergaard J. Beta-adrenergic, cAMP-mediated stimulation of proliferation of brown fat cells in primary culture. Mediation via beta 1 but not via beta 3 adrenoceptors. J Biol Chem. 1992 Jan 25;267(3):2006–2013. [PubMed] [Google Scholar]
  7. Freeman K. B., Heffernan M., Dhalla Z., Patel H. V. Effects of exposure temperature on brown adipose tissue uncoupling protein mRNA levels. Biochem Cell Biol. 1989 Feb-Mar;67(2-3):147–151. doi: 10.1139/o89-022. [DOI] [PubMed] [Google Scholar]
  8. García-Jimenez C., Hernández A., Obregón M. J., Santisteban P. Malic enzyme gene expression in differentiating brown adipocytes: regulation by insulin and triiodothyronine. Endocrinology. 1993 Apr;132(4):1537–1543. doi: 10.1210/endo.132.4.8462451. [DOI] [PubMed] [Google Scholar]
  9. Hargrove J. L., Schmidt F. H. The role of mRNA and protein stability in gene expression. FASEB J. 1989 Oct;3(12):2360–2370. doi: 10.1096/fasebj.3.12.2676679. [DOI] [PubMed] [Google Scholar]
  10. Hentze M. W. Determinants and regulation of cytoplasmic mRNA stability in eukaryotic cells. Biochim Biophys Acta. 1991 Nov 11;1090(3):281–292. doi: 10.1016/0167-4781(91)90191-n. [DOI] [PubMed] [Google Scholar]
  11. Herron D., Rehnmark S., Néchad M., Loncar D., Cannon B., Nedergaard J. Norepinephrine-induced synthesis of the uncoupling protein thermogenin (UCP) and its mitochondrial targeting in brown adipocytes differentiated in culture. FEBS Lett. 1990 Jul 30;268(1):296–300. doi: 10.1016/0014-5793(90)81031-i. [DOI] [PubMed] [Google Scholar]
  12. Jacobsson A., Cannon B., Nedergaard J. Physiological activation of brown adipose tissue destabilizes thermogenin mRNA. FEBS Lett. 1987 Nov 30;224(2):353–356. doi: 10.1016/0014-5793(87)80483-0. [DOI] [PubMed] [Google Scholar]
  13. Jacobsson A., Nedergaard J., Cannon B. alpha- and beta-adrenergic control of thermogenin mRNA expression in brown adipose tissue. Biosci Rep. 1986 Jul;6(7):621–631. doi: 10.1007/BF01114756. [DOI] [PubMed] [Google Scholar]
  14. Jacobsson A., Stadler U., Glotzer M. A., Kozak L. P. Mitochondrial uncoupling protein from mouse brown fat. Molecular cloning, genetic mapping, and mRNA expression. J Biol Chem. 1985 Dec 25;260(30):16250–16254. [PubMed] [Google Scholar]
  15. Kaufmann S. H. Antagonism between camptothecin and topoisomerase II-directed chemotherapeutic agents in a human leukemia cell line. Cancer Res. 1991 Feb 15;51(4):1129–1136. [PubMed] [Google Scholar]
  16. Klaus S., Casteilla L., Bouillaud F., Ricquier D. The uncoupling protein UCP: a membraneous mitochondrial ion carrier exclusively expressed in brown adipose tissue. Int J Biochem. 1991;23(9):791–801. doi: 10.1016/0020-711x(91)90062-r. [DOI] [PubMed] [Google Scholar]
  17. Kohno K., Hamanaka R., Abe T., Nomura Y., Morimoto A., Izumi H., Shimizu K., Ono M., Kuwano M. Morphological change and destabilization of beta-actin mRNA by tumor necrosis factor in human microvascular endothelial cells. Exp Cell Res. 1993 Oct;208(2):498–503. doi: 10.1006/excr.1993.1272. [DOI] [PubMed] [Google Scholar]
  18. Mitchell J. R., Jacobsson A., Kirchgessner T. G., Schotz M. C., Cannon B., Nedergaard J. Regulation of expression of the lipoprotein lipase gene in brown adipose tissue. Am J Physiol. 1992 Sep;263(3 Pt 1):E500–E506. doi: 10.1152/ajpendo.1992.263.3.E500. [DOI] [PubMed] [Google Scholar]
  19. Müllner E. W., Kühn L. C. A stem-loop in the 3' untranslated region mediates iron-dependent regulation of transferrin receptor mRNA stability in the cytoplasm. Cell. 1988 Jun 3;53(5):815–825. doi: 10.1016/0092-8674(88)90098-0. [DOI] [PubMed] [Google Scholar]
  20. Nedergaard J., Lindberg O. The brown fat cell. Int Rev Cytol. 1982;74:187–286. doi: 10.1016/s0074-7696(08)61173-0. [DOI] [PubMed] [Google Scholar]
  21. Nielsen D. A., Shapiro D. J. Insights into hormonal control of messenger RNA stability. Mol Endocrinol. 1990 Jul;4(7):953–957. doi: 10.1210/mend-4-7-953. [DOI] [PubMed] [Google Scholar]
  22. Niijima A., Rohner-Jeanrenaud F., Jeanrenaud B. Role of ventromedial hypothalamus on sympathetic efferents of brown adipose tissue. Am J Physiol. 1984 Oct;247(4 Pt 2):R650–R654. doi: 10.1152/ajpregu.1984.247.4.R650. [DOI] [PubMed] [Google Scholar]
  23. Néchad M., Nedergaard J., Cannon B. Noradrenergic stimulation of mitochondriogenesis in brown adipocytes differentiating in culture. Am J Physiol. 1987 Dec;253(6 Pt 1):C889–C894. doi: 10.1152/ajpcell.1987.253.6.C889. [DOI] [PubMed] [Google Scholar]
  24. Obregón M. J., Jacobsson A., Kirchgessner T., Schotz M. C., Cannon B., Nedergaard J. Postnatal recruitment of brown adipose tissue is induced by the cold stress experienced by the pups. An analysis of mRNA levels for thermogenin and lipoprotein lipase. Biochem J. 1989 Apr 15;259(2):341–346. doi: 10.1042/bj2590341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Patel H. V., Freeman K. B., Desautels M. Selective loss of uncoupling protein mRNA in brown adipose tissue on deacclimation of cold-acclimated mice. Biochem Cell Biol. 1987 Nov;65(11):955–959. doi: 10.1139/o87-124. [DOI] [PubMed] [Google Scholar]
  26. Peltz S. W., Jacobson A. mRNA stability: in trans-it. Curr Opin Cell Biol. 1992 Dec;4(6):979–983. doi: 10.1016/0955-0674(92)90129-z. [DOI] [PubMed] [Google Scholar]
  27. Puigserver P., Herron D., Gianotti M., Palou A., Cannon B., Nedergaard J. Induction and degradation of the uncoupling protein thermogenin in brown adipocytes in vitro and in vivo. Evidence for a rapidly degradable pool. Biochem J. 1992 Jun 1;284(Pt 2):393–398. doi: 10.1042/bj2840393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rehnmark S., Antonson P., Xanthopoulos K. G., Jacobsson A. Differential adrenergic regulation of C/EBP alpha and C/EBP beta in brown adipose tissue. FEBS Lett. 1993 Mar 8;318(3):235–241. doi: 10.1016/0014-5793(93)80519-z. [DOI] [PubMed] [Google Scholar]
  29. Rehnmark S., Bianco A. C., Kieffer J. D., Silva J. E. Transcriptional and posttranscriptional mechanisms in uncoupling protein mRNA response to cold. Am J Physiol. 1992 Jan;262(1 Pt 1):E58–E67. doi: 10.1152/ajpendo.1992.262.1.E58. [DOI] [PubMed] [Google Scholar]
  30. Rehnmark S., Kopecký J., Jacobsson A., Néchad M., Herron D., Nelson B. D., Obregon M. J., Nedergaard J., Cannon B. Brown adipocytes differentiated in vitro can express the gene for the uncoupling protein thermogenin: effects of hypothyroidism and norepinephrine. Exp Cell Res. 1989 May;182(1):75–83. doi: 10.1016/0014-4827(89)90280-2. [DOI] [PubMed] [Google Scholar]
  31. Rehnmark S., Néchad M., Herron D., Cannon B., Nedergaard J. Alpha- and beta-adrenergic induction of the expression of the uncoupling protein thermogenin in brown adipocytes differentiated in culture. J Biol Chem. 1990 Sep 25;265(27):16464–16471. [PubMed] [Google Scholar]
  32. Reichling S., Ridley R. G., Patel H. V., Harley C. B., Freeman K. B. Loss of brown adipose tissue uncoupling protein mRNA on deacclimation of cold-exposed rats. Biochem Biophys Res Commun. 1987 Feb 13;142(3):696–701. doi: 10.1016/0006-291x(87)91470-7. [DOI] [PubMed] [Google Scholar]
  33. Ricquier D., Bouillaud F., Toumelin P., Mory G., Bazin R., Arch J., Pénicaud L. Expression of uncoupling protein mRNA in thermogenic or weakly thermogenic brown adipose tissue. Evidence for a rapid beta-adrenoreceptor-mediated and transcriptionally regulated step during activation of thermogenesis. J Biol Chem. 1986 Oct 25;261(30):13905–13910. [PubMed] [Google Scholar]
  34. Ricquier D., Mory G., Bouillaud F., Thibault J., Weissenbach J. Rapid increase of mitochondrial uncoupling protein and its mRNA in stimulated brown adipose tissue. Use of a cDNA probe. FEBS Lett. 1984 Dec 10;178(2):240–244. doi: 10.1016/0014-5793(84)80608-0. [DOI] [PubMed] [Google Scholar]
  35. Sachs A. The role of poly(A) in the translation and stability of mRNA. Curr Opin Cell Biol. 1990 Dec;2(6):1092–1098. doi: 10.1016/0955-0674(90)90161-7. [DOI] [PubMed] [Google Scholar]
  36. Schwartz M. L., Shneidman P. S., Bruce J., Schlaepfer W. W. Actinomycin prevents the destabilization of neurofilament mRNA in primary sensory neurons. J Biol Chem. 1992 Dec 5;267(34):24596–24600. [PubMed] [Google Scholar]
  37. Shapiro D. J., Blume J. E., Nielsen D. A. Regulation of messenger RNA stability in eukaryotic cells. Bioessays. 1987 May;6(5):221–226. doi: 10.1002/bies.950060507. [DOI] [PubMed] [Google Scholar]
  38. Silva J. E. Full expression of uncoupling protein gene requires the concurrence of norepinephrine and triiodothyronine. Mol Endocrinol. 1988 Aug;2(8):706–713. doi: 10.1210/mend-2-8-706. [DOI] [PubMed] [Google Scholar]
  39. Tvrdík P., Kuzela S., Houstek J. Low translational efficiency of the F1-ATPase beta-subunit mRNA largely accounts for the decreased ATPase content in brown adipose tissue mitochondria. FEBS Lett. 1992 Nov 16;313(1):23–26. doi: 10.1016/0014-5793(92)81175-l. [DOI] [PubMed] [Google Scholar]
  40. Young J. B., Saville E., Rothwell N. J., Stock M. J., Landsberg L. Effect of diet and cold exposure on norepinephrine turnover in brown adipose tissue of the rat. J Clin Invest. 1982 May;69(5):1061–1071. doi: 10.1172/JCI110541. [DOI] [PMC free article] [PubMed] [Google Scholar]

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