The Korean Journal of Physiology and Pharmacology

The Korean Society of Pharmacology (대한약리학회)
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Effect of Modulation of hnRNP L Levels on the Decay of bcl-2 mRNA in MCF-7 Cells

  • Lim, Mi-Hyun (Department of Biochemistry, College of Medicine, The Catholic University of Korea) ;
  • Lee, Dong-Hyoung (Department of Biochemistry, College of Medicine, The Catholic University of Korea) ;
  • Jung, Seung-Eun (Department of Biochemistry, College of Medicine, The Catholic University of Korea) ;
  • Youn, Dong-Ye (Department of Biochemistry, College of Medicine, The Catholic University of Korea) ;
  • Park, Chan-Sun (Bioindustry Research Center, Korea Research Institute Bioscience and Biotechnology) ;
  • Lee, Jeong-Hwa (Department of Biochemistry, College of Medicine, The Catholic University of Korea)
  • Published : 2010.02.28
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Abstract

It has been shown that CA repeats in the 3'-untranslated region (UTR) of bcl-2 mRNA contribute the constitutive decay of bcl-2 mRNA and that hnRNP L (heterogenous nuclear ribonucleoprotein L) interacts with CA repeats in the 3'-UTR of bcl-2 mRNA, both in vitro and in vivo. The aim of this study was to determine whether the alteration of hnRNP L affects the stability of bcl-2 mRNA in vivo. Human breast carcinoma MCF-7 cells were transfected with hnRNP L-specific shRNA or hnRNP L-expressing vector to decrease or increase hnRNP L levels, respectively, followed by an actinomycin D chase. An RT-PCR analysis showed that the rate of degradation of endogenous bcl-2 mRNA was not affected by the decrease or increase in the hnRNP L levels. Furthermore, during apoptosis or autophagy, in which bcl-2 expression has been reported to decrease, no difference in the degradation of bcl-2 mRNA was observed between control and hnRNP L-knock down MCF-7 Cells. On the other hand, the levels of AUF-1 and nucleolin, transacting factors for ARE in the 3'UTR of bcl-2 mRNA, were not significantly affected by the decrease in hnRNP L, suggesting that a disturbance in the quantitative balance between these transacting factors is not likely to interfere with the effect of hnRNP L. Collectively, the findings indicate that the decay of bcl-2 mRNA does not appear to be directly controlled by hnRNP L in vivo.

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References

  1. Heath-Engel HM, Chang NC, Shore GC. The endoplasmic reticulum in apoptosis and autophagy: role of the BCL-2 protein family. Oncogene. 2008;27:6419-6433. https://doi.org/10.1038/onc.2008.309
  2. Reed JC. Bcl-2 and the regulation of programmed cell death. J Cell Biol 1994;124:1-6. https://doi.org/10.1083/jcb.124.1.1
  3. Davies AM. The Bcl-2 family of proteins, and the regulation of neuronal survival. Trends Neurosci. 1995;18:355-358. https://doi.org/10.1016/0166-2236(95)93928-Q
  4. Greenlund LJ, Korsmeyer SJ, Johnson EM Jr. Role of BCL-2 in the survival and function of developing and mature sympathetic neurons. Neuron. 1995;15:649-661. https://doi.org/10.1016/0896-6273(95)90153-1
  5. Young RL, Korsmeyer SJ. A negative regulatory element in the bcl-2 5'-untranslated region inhibits expression from an upstream promoter. Mol Cell Biol. 1993;13:3686-3697. https://doi.org/10.1128/MCB.13.6.3686
  6. Perillo B, Sasso A, Abbondanza C, Palumbo G. 17beta-estradiol inhibits apoptosis in MCF-7 cells, inducing bcl-2 expression via two estrogen-responsive elements present in the coding sequence. Mol Cell Biol. 2000;20:2890-2901. https://doi.org/10.1128/MCB.20.8.2890-2901.2000
  7. Riordan FA, Foroni L, Hoffbrand AV, Mehta AB, Wickremasinghe RG. Okadaic acid-induced apoptosis of HL60 leukemia cells is preceded by destabilization of bcl-2 mRNA and down-regulation of bcl-2 protein. FEBS Lett. 1998;435:195-198. https://doi.org/10.1016/S0014-5793(98)01070-9
  8. Bandyopadhyay S, Sengupta TK, Fernandes DJ, Spicer EK. Taxol- and okadaic acid-induced destabilization of bcl-2 mRNA is associated with decreased binding of proteins to a bcl-2 instability element. Biochem Pharmacol 2003;66:1151-1162. https://doi.org/10.1016/S0006-2952(03)00453-2
  9. Guhaniyogi J, Brewer G. Regulation of mRNA stability in mammalian cells. Gene. 2001;265:11-23. https://doi.org/10.1016/S0378-1119(01)00350-X
  10. Schiavone N, Rosini P, Quattrone A, Donnini M, Lapucci A, Citti L, Bevilacqua A, Nicolin A, Capaccioli S. A conserved AU-rich element in the 3' untranslated region of bcl-2 mRNA is endowed with a destabilizing function that is involved in bcl-2 down-regulation during apoptosis. FASEB J. 2000;14:174-184. https://doi.org/10.1096/fasebj.14.1.174
  11. Donnini M, Lapucci A, Papucci L, Witort E, Tempestini A, Brewer G, Bevilacqua A, Nicolin A, Capaccioli S, Schiavone N. Apoptosis is associated with modifications of bcl-2 mRNA AU-binding proteins. Biochem Biophys Res Commun. 2001;287:1063-1069. https://doi.org/10.1006/bbrc.2001.5700
  12. Lapucci A, Donnini M, Papucci L, Witort E, Tempestini A, Bevilacqua A, Nicolin A, Brewer G, Schiavone N, Capaccioli S. AUF-1 Is a bcl-2 A + U-rich element-binding protein involved in bcl-2 mRNA destabilization during apoptosis. J Biol Chem. 2002;277:16139-16146. https://doi.org/10.1074/jbc.M201377200
  13. Sengupta TK, Bandyopadhyay S, Fernandes DJ, Spicer EK. Identification of nucleolin as an AU-rich element binding protein involved in bcl-2 mRNA stabilization. J Biol Chem. 2004;279:10855-10863. https://doi.org/10.1074/jbc.M309111200
  14. Donnini M, Lapucci A, Papucci L, Witort E, Jacquier A, Brewer G, Nicolin A, Capaccioli S, Schiavone N. Identification of TINO: a new evolutionarily conserved BCL-2 AU-rich element RNA-binding protein. J Biol Chem. 2004;279:20154-20166. https://doi.org/10.1074/jbc.M314071200
  15. Lee JH, Jeon MH, Seo YJ, Lee YJ, Ko JH, Tsujimoto Y, Lee JH. CA repeats in the 3'-untranslated region of bcl-2 mRNA mediate constitutive decay of bcl-2 mRNA. J Biol Chem. 2004;279:42758-42764. https://doi.org/10.1074/jbc.M407357200
  16. Lee DH, Lim MH, Youn DY, Jung SE, Ahn YS, Tsujimoto Y, Lee JH. hnRNP L binds to CA repeats in the 3'UTR of bcl-2 mRNA. Biochem Biophys Res Commun. 2009;382:583-587. https://doi.org/10.1016/j.bbrc.2009.03.069
  17. Park HG, Yoon JY, Choi M. Heterogeneous nuclear ribonucleoprotein D/AUF1 interacts with heterogeneous nuclear ribonucleoprotein L. J Biosci. 2007;32:1263-1272. https://doi.org/10.1007/s12038-007-0135-8
  18. Miyashita T, Krajewski S, Krajewska M, Wang HG, Lin HK, Liebermann DA, Hoffman B, Reed JC. Tumor suppressor p53 is a regulator of bcl-2 and bax gene expression in vitro and in vivo. Oncogene. 1994;9:1799-1805.
  19. Suzuki A, Matsuzawa A, Iguchi T. Down regulation of Bcl-2 is the first step on Fas-mediated apoptosis of male reproductive tract. Oncogene. 1996;13:31-37.
  20. Pattingre S, Tassa A, Qu X, Garuti R, Liang XH, Mizushima N, Packer M, Schneider MD, Levine B. Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy. Cell. 2005;122:927-939. https://doi.org/10.1016/j.cell.2005.07.002
  21. Hui J, Reither G, Bindereif A. Novel functional role of CA repeats and hnRNP L in RNA stability. RNA. 2003;9:931-936. https://doi.org/10.1261/rna.5660803
  22. Hui J, Hung LH, Heiner M, Schreiner S, Neumuller N, Reither G, Haas SA, Bindereif A. Intronic CA-repeat and CA-rich elements: a new class of regulators of mammalian alternative splicing. EMBO J. 2005;24:1988-1998. https://doi.org/10.1038/sj.emboj.7600677
  23. Rothrock CR, House AE, Lynch KW. HnRNP L represses exon splicing via a regulated exonic splicing silencer. EMBO J. 2005;24:2792-2802. https://doi.org/10.1038/sj.emboj.7600745
  24. Shih SC, Claffey KP. Regulation of human vascular endothelial growth factor mRNA stability in hypoxia by heterogeneous nuclear ribonucleoprotein L. J Biol Chem. 1995;274:1359-1365.
  25. Soderberg M, Raffalli-Mathieu F, Lang MA. Identification of a regulatory cis-element within the 3'-untranslated region of the murine inducible nitric oxide synthase (iNOS) mRNA; interaction with heterogeneous nuclear ribonucleoproteins I and L and role in the iNOS gene expression. Mol Immunol. 2007;44:434-442. https://doi.org/10.1016/j.molimm.2006.02.019
  26. Hamilton BJ, Nichols RC, Tsukamoto H, Boado RJ, Pardridge WM, Rigby WF. hnRNP A2 and hnRNP L bind the 3'UTR of glucose transporter 1 mRNA and exist as a complex in vivo. Biochem Biophys Res Commun. 1999;261:646-651. https://doi.org/10.1006/bbrc.1999.1040

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