Abstract
Expression of the prion protein is necessary for infection with prion diseases. Altered expression levels may play an important role in susceptibility to infection. Therefore, understanding the mechanisms that regulate prion protein expression is of great importance. It was previously shown that expression of the prion protein is to some degree regulated by an alternative promoter within intron 1. Studies using GFP and luciferase reporter systems were undertaken to determine key sites for the repression and activation of expression of the prion protein driven by intron 1. We identified a region within intron 1 sufficient to drive prion protein expression. Our findings highlight two potential repressor regions. Both regions have binding sites for the known repressor Hes-1. Hes-1 overexpression caused a dramatic decrease in PrP protein expression. Additionally, we have identified Atox-1 as a transcription factor that upregulates prion protein expression. These findings clearly indicate that intron 1 plays a key role in regulation of prion protein expression levels.
Similar content being viewed by others
References
Prusiner SB (1998) Prions. Proc Natl Acad Sci USA 95:13363–13383
Büeler H, Aguzzi A, Sailer A, Greiner RA, Autenried P, Aguet M, Weissmann C (1993) Mice devoid of PrP are resistant to scrapie. Cell 73:1339–1347
Mallucci G, Dickinson A, Linehan J, Klöhn P-C, Brandner S, Collinge J (2003) Depleting neuronal PrP in prion infection prevents disease and reverses spongiosis. Science 302:871–874
Fischer M, Rulicke T, Raeber A, Sailer A, Moser M, Oesch B, Brandner S, Aguzzi A, Weissmann C (1996) Prion protein (PrP) with amino-proximal deletions restoring susceptibility of PrP knockout mice to scrapie. EMBO J 15:1255–1264
Li G, Bolton D (1997) A novel hamster prion protein mRNA contains an extra exon: increased expression in scrapie. Brain Res 751:265–274
Lee IY, Westaway D, Smit AFA, Wang K, Seto J, Chen L, Acharya C, Ankener M, Baskin D, Cooper C, Yao H, Prusiner SB, Hood LE (1998) Complete genomic sequence and analysis of the prion protein gene region from three mammalian species. Genome Res 8:1022–1037
Haigh CL, Wright JA, Brown DR (2007) Regulation of prion protein expression by non coding regions of the Prnp gene. J Mol Biol 368:915–927
Inoue S, Tanaka M, Horiuchi M, Ishiguro N, Shinagawa M (1997) Characterisation of the bovine prion protein gene: the expression requires interaction between the promoter and intron. J Vet Med Sci 59:175–183
Bellingham SA, Coleman LA, Masters CL, Camakaris J, Hill AF (2009) Regulation of prion gene expression by transcription factors SP1 and metal transcription factor-1. J Biol Chem 284:1291–1301
Burgess ST, Shen C, Ferguson LA, O’Neill GT, Docherty K, Hunter N, Goldmann W (2009) Identification of adjacent binding sites for the YY1 and E4BP4 transcription factors in the ovine PrP (Prion) gene promoter. J Biol Chem 284:6716–6724
Shyu WC, Kao MC, Chou WY, Hsu YD, Soong BW (2000) Heat shock modulates prion protein expression in human NT-2 cells. Mol Neurosci 11:771–774
Shyu WC, Harn HJ, Saeki K, Kubosaki A, Matsumoto Y, Onodera T, Chen CJ, Hsu YD, Chiang YH (2002) Molecular modulation of expression of prion protein by heat shock. Mol Neurobiol 26:1–12
Wang V, Chuang TC, Hsu YD, Chou WY, Kao MC (2005) Nitric oxide induces prion protein via MEK and p38 MAPK signalling. Biochem Biophys Res Comm 333:95–100
Shyu WC, Lin SZ, Saeki K, Kubosaki A, Matsumoto Y, Onodera T, Chiang MF, Thajeb P, Li H (2004) Hyperbaric oxygen enhances the expression of prion protein and heat shock protein 70 in a mouse neuroblastoma cell line. Cell Mol Neurobiol 24:257–268
Shyu WC, Chen CP, Saeki K, Kubosaki A, Matsumoto Y, Onodera T, Ding DC, Chiang MF, Lee YJ, Lin SZ, Li H (2005) Hypoglycaemia enhances the expression of prion protein and heat-shock protein 70 in a mouse neuroblastoma cell line. J Neurosci Res 80:887–894
Qin K, Zhao L, Ash RD, McDonough WF, Zhao RY (2009) ATM-mediated transcriptional elevation of prion in response to copper-induced oxidative stress. J Biol Chem 284:4582–4593
Rybner C, Hilion J, Sahraoui T, Lanotte M, Botti J (2002) All-trans retinoic acid down-regulates prion protein expression independently of granulocyte maturation. Leukaemia 16:940–948
Holme A, Daniels M, Sassoon J, Brown DR (2003) A novel method of generating neuronal cell lines from gene-knockout mice to study prion protein membrane orientation. Eur J Neurosci 18:571–579
Lenhard B, Sandelin A, Mendoza L, Engström P, Jareborg N, Wasserman WW (2003) Identification of conserved regulatory elements by comparative genome analysis. J Biol 2:13
Brown DR, Schmidt B, Kretzschmar HA (1997) Expression of prion protein in PC12 is enhanced by exposure to oxidative stress. Int J Dev Neurosci 15:961–972
Ishii T, Itoh K, Takahashi S, Sato H, Yanagawa T, Katoh Y, Bannai S, Yamamoto M (2000) Transcription factor Nrf2 coordinately regulates a group of oxidative stress-inducible genes in macrophages. J Biol Chem 275:16023–16029
Radtke F, Heuchel R, Georgiev O, Hergersberg M, Gariglio M, Dembic Z, Schaffner W (1993) Cloned transcription factor MTF-1 activates the mouse metallothionein I promoter. EMBO J 12:1355–1362
Itoh S, Kim HW, Nakagawa O, Ozumi K, Lessner SM, Aoki H, Akram K, McKinney RD, Ushio-Fukai M, Fukai T (2008) Novel role of antioxidant-1 (Atox1) as a copper-dependent transcription factor involved in cell proliferation. J Biol Chem 283:9157–9167
Kotomura N, Ninomiya Y, Umesono K, Niwa O (1997) Transcriptional regulation by competition between ELP isoforms and nuclear receptors. Biochem Biophys Res Comm 230:407–412
Imbriano C, Gurtner A, Cocchiarella F, Di Agostino S, Basile V, Gostissa M, Dobbelstein M, Del Sal G, Piaggio G, Mantovani R (2005) Direct p53 transcriptional repression: in vivo analysis of CCAAT-containing G2/M promoters. Mol Cell Biol 25:3737–3751
Gaubatz S, Imhof A, Dosch R, Werner O, Mitchell P, Buettner R, Eilers M (1995) Transcriptional activation by Myc is under negative control by the transcription factor AP-2. EMBO J 14:1508–1519
Takebayashi K, Sasai Y, Sakai Y, Watanabe T, Nakanishi S, Kageyama R (1994) Structure, chromosomal locus, and promoter analysis of the gene encoding the mouse helix-loop-helix factor HES-1. Negative autoregulation through the multiple N box elements. J Biol Chem 269:5150–5156
Funke-Kaiser H, Theis S, Behrouzi T, Thomas A, Scheuch K, Zollmann FS, Paterka M, Paul M, Orzechowski HD (2001) Functional characterization of the human prion protein promoter in neuronal and endothelial cells. J Mol Med 79:529–535
Mahal SP, Asante EA, Antoniou M, Collinge J (2001) Isolation and functional characterisation of the promoter region of the human prion protein gene. Gene 268:105–114
Brown DR (2004) Role of the prion protein in copper turnover in astrocytes. Neurobiol Dis 15:534–543
Armendariz AD, Gonzalez M, Loguinov AV, Vulpe CD (2004) Gene expression profiling in chronic copper overload reveals upregulation of Prnp and APP. Physiol Genomics 20:45–54
Varela-Nallar L, Toledo EM, Larrondo LF, Cabral AL, Martins VR, Inestrosa NC (2006) Induction of cellular prion protein gene expression by copper in neurons. Am J Physiol Cell Physiol 290:C271–C281
Naeve GS, Vana AM, Eggold JR, Kelner GS, Maki R, Desouza EB, Foster AC (1999) Expression profile of the copper homeostasis gene, rAtox1, in the rat brain. Neuroscience 93:1179–1187
Muller PA, Klomp LW (2009) ATOX1: a novel copper-responsive transcription factor in mammals? Int J Biochem Cell Biol 41:1233–1236
Itoh S, Ozumi K, Kim HW, Nakagawa O, McKinney RD, Folz RJ, Zelko IN, Ushio-Fukai M, Fukai T (2009) Novel mechanism for regulation of extracellular SOD transcription and activity by copper: role of antioxidant-1. Free Radic Biol Med 46:95–104
Kralovicova S, Fontaine SN, Alderton A, Alderman J, Ragnarsdottir KV, Collins SJ, Brown DR (2009) The effects of prion protein expression on metal metabolism. Mol Cell Neurosci 41:135–147
McCormack JE, Baybutt HN, Everington D, Will RG, Ironside JW, Manson JC (2002) PRNP contains both intronic and upstream regulatory regions that may influence susceptibility to Creutzfeldt-Jakob disease. Gene 288:139–146
Seabury CM, Womack JE, Piedrahita J, Derr JN (2004) Comparative PRNP genotyping of US cattle sires for potential association with BSE. Mamm Genome 15:828–833
Nakamitsu S, Miyazawa T, Horiuchi M, Onoe S, Ohoba Y, Kitagawa H, Ishiguro N (2006) Sequence variation of bovine prion protein gene in Japanese cattle (Holstein and Japanese Black). J Vet Med Sci 68:27–33
Kerber AR, Hepp D, Passos DT, de Azevedo Weimer T (2008) Polymorphisms of two indels at the PRNP gene in three beef cattle herds. Biochem Genet 46:1–7
Li R, Liu T, Wong B-S, Pan T, Morillas M, Swietnicki W, O'Rourke K et al (2000) Identification of an epitope in the C terminus of normal prion protein whose expression is modulated by binding events in the N terminus. J Mol Biol 301:567–573
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wright, J.A., McHugh, P.C., Stockbridge, M. et al. Activation and repression of prion protein expression by key regions of intron 1. Cell. Mol. Life Sci. 66, 3809–3820 (2009). https://doi.org/10.1007/s00018-009-0154-8
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00018-009-0154-8