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. 2004 Dec;24(6):793–814. doi: 10.1007/s10571-004-6920-0

Gene Expression Profile Following Stable Expression of the Cellular Prion Protein

Jun-ichi Satoh 1,✉, Takashi Yamamura 1
PMCID: PMC11529964  PMID: 15672681

Abstract

1. The cellular prion protein (PrPC) is expressed widely in neural and nonneural tissues at the highest level in neurons in the central nervous system (CNS).

2. Recent studies indicated that transgenic mice with the cytoplasmic accumulation of PrPC exhibited extensive neurodegeneration in the cerebellum, although the underlying mechanism remains unknown. To identify the genes whose expression is controlled by overexpression of PrPC in human cells, we have established a stable PrPC-expressing HEK293 cell line designated P1 by the site-specific recombination technique.

3. Microarray analysis identified 33 genes expressed differentially between P1 and the parent PrPC-non-expressing cell line among 12,814 genes examined. They included 18 genes involved in neuronal and glial functions, 5 related to production of extracellular matrix proteins, and 2 located in the complement cascade.

4. Northern blot analysis verified marked upregulation in P1 of the brain-specific protein phosphatase 2A beta subunit (PPP2R2B), a causative gene of spinocerebellar ataxia 12, and the cerebellar degeneration-related autoantigen (CDR34) gene associated with development of paraneoplastic cerebellar degeneration.

5. These results indicate that accumulation of PrPC in the cell caused aberrant regulation of a battery of the genes important for specific neuronal function. This represents a possible mechanism underlying PrPC-mediated selective neurodegeneration.

Keywords: CDR34, cellular prion protein, complement, extracellular matrix, microarray, neurodegeneration, PPP2R2B.

References

  1. Bendheim, P. E., Brown, H. R., Rudelli, R. D., Scala, L. J., Goller, N. L., Wen, G. Y., Kascsak, R. J., Cashman, N. R., and Bolton, D. C. (1992). Nearly ubiquitous tissue distribution of the scrapie agent precursor protein. Neurology42:149–156. [DOI] [PubMed] [Google Scholar]
  2. Benndorf, R., Sun, X., Gilmont, R. R., Biederman, K. J., Molloy, M. P., Goodmurphy, C. W., Cheng, H., Andrews, P. C., and Welsh, M. J. (2001). HSP22, a new member of the small heat shock protein superfamily, interacts with mimic of phosphorylated HSP27 (3DHSP27). J. Biol. Chem.276:26753–26761. [DOI] [PubMed] [Google Scholar]
  3. Berglund, E. O., Murai, K. K., Fredette, B., Sekerková, G., Marturano, B., Weber, L., Mugnaini, E., and Ranscht, B. (1999). Ataxia and abnormal cerebellar microorganization in mice with ablated contactin gene expression. Neuron24:739–750. [DOI] [PubMed] [Google Scholar]
  4. Brown, D. R., Qin, K., Herms, J. W., Madlung, A., Manson, J., Strome, R., Fraser, P. E., Kruck, T., von Bohlen, A., Schulz-Schaeffer, W., Giese, A., Westaway, D., and Kretzschmar, H. (1997). The cellular prion protein binds copper in vivo. Nature390:684–687. [DOI] [PubMed] [Google Scholar]
  5. Büeler, H., Fischer, M., Lang, Y., Bluethmann, H., Lipp, H.-P., DeArmond, S. J., Prusiner, S. B., Aguet, M., and Weissmann, C. (1992). Normal development and behaviour of mice lacking the neuronal cell-surface PrP protein. Nature356:577–582. [DOI] [PubMed] [Google Scholar]
  6. Camby, I., Belot, N., Lefranc, F., Sadeghi, N., de Launoit, Y., Kaltner, H., Musette, S., Darro, F., Danguy, A., Salmon, I., Gabius, H.-J., and Kiss, R. (2002). Galectin-1 modulates human glioblastoma cell migration into the brain through modifications to the actin cytoskeleton and levels of expression of small GTPases. J. Neuropathol. Exp. Neurol.61:585–596. [DOI] [PubMed] [Google Scholar]
  7. Chen, Y.-T., Rettig, W., Yenamandra, A. K., Kozak, C. A., Chaganti, R. S. K., Poser, J. B., and Old, L. J. (1990). Cerebellar degeneration-related antigen: A highly conserved neuroectodermal marker mapped to chromosomes X in human and mouse. Proc. Natl. Acad. Sci. U.S.A.87:3077–3081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chiarini, L. B., Freitas, A. R. O., Zanata, S. M., Brentani, R. R., Martins, V. R., and Linden, R. (2002). Cellular prion protein transduces neuroprotective signals. EMBO J.21:3317–3326. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Collinge, J., Whittington, M. A., Sidle, K. C. L., Smith, C. J., Palmer, M. S., Clarke, A. R., and Jefferys, J. G. R. (1994). Prion protein is necessary for normal synaptic function. Nature370:295–297. [DOI] [PubMed] [Google Scholar]
  10. DeArmond, S. J., Sánchez, H., Yehiely, F., Qiu, Y., Ninchak-Casey, A., Daggett, V., Camerino, A. P., Cayetano, J., Rogers, M., Groth, D., Torchia, M., Tremblay, P., Scott, M. R., Cohen, F. E., and Prusiner, S. B. (1997). Selective neuronal targeting in prion disease. Neuron19:1337–1348. [DOI] [PubMed] [Google Scholar]
  11. Dimcheff, D. E., Portis, J. L., and Caughey, B. (2003). Prion proteins meet protein quality control. Trends Cell Biol.13:337–340. [DOI] [PubMed] [Google Scholar]
  12. Finch, P. W., He, X., Kelley, M. J., Üren, A., Schaudies, R. P., Popescu, N. C., Rudikoff, S., Aaronson, S. A., Varmus, H. E., and Rubin, J. S. (1997). Purification and molecular cloning of a secreted, frizzled-related antagonist of Wnt action. Proc. Natl. Acad. Sci. U.S.A.94:6770–6775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fujigasaki, H., Verma, I. C., Camuzat, A., Margolis, R. L., Zander, C., Lebre, A.-S., Jamot, L., Saxena, R., Anand, I., Holmes, S. E., Ross, C. A., Dürr, A., and Brice, A. (2001). SCA12 is a rare locus for autosomal dominant cerebellar ataxia: A study of an Indian family. Ann. Neurol.49:117–121. [PubMed] [Google Scholar]
  14. Graner, E., Mercadante, A. F., Zanata, S. M., Forlenza, O. V., Cabral, A. L. B., Veiga, S. S., Juliano, M. A., Roesler, R., Walz, R., Minetti, A., Izquierdo, I., Martins, V. R., and Brentani, R. R. (2000). Cellular prion protein binds laminin and mediates neuritogenesis. Mol. Brain Res.76:85–92. [DOI] [PubMed] [Google Scholar]
  15. Herms, J. W., Tings, T., Dunker, S., and Kretzschmar, H. A. (2001). Prion protein affects Ca$^{2+}$-activated K$^{+}$ currents in cerebellar Purkinje cells. Neurobiol. Dis.8:324–330. [DOI] [PubMed] [Google Scholar]
  16. Holmes, S. E., O’Hearn, E. E., McInnis, M. G., Gorelick-Feldman, D. A., Kleiderlein, J. J., Callahan, C., Kwak, N. G., Ingersoll-Ashworth, R. G., Sherr, M., Sumner, A. J., Sharp, A. H., Ananth, U., Seltzer, W. K., Boss, M. A., Vieria-Saecker, A.-M., Epplen, J. T., Riess, O., Ross, C. A., and Margolis, R. L. (1999). Expansion of a novel CAG trinucleotide repeat in the 5′ region of PPP2R2B is associated with SCA12. Nature Genet.23:391–392. [DOI] [PubMed] [Google Scholar]
  17. Hosoda, K., Hammer, R. E., Richardson, J. A., Baynash, A. G., Cheung, J. C., Giaid, A., and Yanagisawa, M. (1994). Targeted and natural (piebald-lethal) mutations of endothelin-B receptor gene produce megacolon associated with spotted coat color in mice. Cell79:1267–1276. [DOI] [PubMed] [Google Scholar]
  18. Hundt, C., Peyrin, J.-M., Haík, S., Gauczynski, S., Leucht, C., Rieger, R., Riley, M. L., Deslys, J.-P., Dormont, D., Lasmézas, C. I., and Weiss, S. (2001). Identification of interaction domains of the prion protein with its 37-kDa/67-kda laminin receptor. EMBO J.20:5876–5886. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Inagaki, N., Tsuura, Y., Namba, N., Masuda, K., Gonoi, T., Horie, M., Seino, Y., Mizuta, M., and Seino, S. (1995). Cloning and functional characterization of a novel ATP-sensitive potassium channel ubiquitously expressed in rat tissues, including pancreatic islets, pituitary, skeletal muscle, and heart. J. Biol. Chem.270:5691–5694. [DOI] [PubMed] [Google Scholar]
  20. Kawaguchi, S.-Y., and Hirano, T. (2002). Signaling cascade regulating long-term potentiation of GABAA receptor responsiveness in cerebellar Purkinje neurons. J. Neurosci.22:3969–3976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Klein, M. A., Kaeser, P. S., Schwarz, P., Weyd, H., Xenarios, I., Zinnkernagel, R. M., Carroll, M. C., Verbeek, J. S., Botto, M., Walport, M. J., Molina, H., Kalinke, U., Acha-Orbea, H., and Aguzzi, A. (2001). Complement facilitates early prion pathogenesis. Nature Med.7:488–492. [DOI] [PubMed] [Google Scholar]
  22. Koike, F., Satoh, J.-I., Miyake, S., Yamamoto, T., Kawai, M., Kikuchi, S., Nomura, K., Yokoyama, K., Ota, K., Kanda, T., Fukazawa, T., and Yamamura, T. (2003). Microarray analysis identifies interferon β-regulated genes in multiple sclerosis. J. Neuroimmunol.139:109–118. [DOI] [PubMed] [Google Scholar]
  23. Kuwahara, C., Takeuchi, A. M., Nishimura, T., Haraguchi, K., Kubosaki, A., Matsumoto, Y., Saeki, K., Matsumoto, Y., Yokoyama, T., Itohara, S., and Onodera, T. (1999). Prions prevent neural cell-line death. Nature400:225–226. [DOI] [PubMed] [Google Scholar]
  24. Lacroix, M., Ebel, C., Kardos, J., Dobó, J., Gál, P., Závodszky, P., Arlaud, G. J., and Thielens, N. M. (2001). Assembly and enzymatic properties of the catalytic domain of human complement protease C1r. J. Biol. Chem.276:36233–36240. [DOI] [PubMed] [Google Scholar]
  25. Leclerc, P. C., Auger-Messier, M., Lanctot, P. M., Escher, E., Leduc, R., and Guillemette, G. (2002). A polyaromatic caveolin-binding-like motif in the cytoplasmic tail of the type 1 receptor for angiotensin II plays an important role in receptor trafficking and signaling. Endocrinology143:4702– 4710. [DOI] [PubMed] [Google Scholar]
  26. Legname, G., Nelken, P., Guan, Z., Kanyo, Z. F., DeArmond, S. J., and Prusiner, S. B. (2002). Prion and doppel proteins bind to granule cells of the cerebellum. Proc. Natl. Acad. Sci. U.S.A.99:16285– 16290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mabbott, N. A., Bruce, M. E., Botto, M., Walport, M. J., and Pepys, M. B. (2001). Temporary depletion of complement component C3 or genetic deficiency of C1q significantly delays onset of scrapie. Nature Med.7:485–487. [DOI] [PubMed] [Google Scholar]
  28. Ma, J., Wollmann, R., and Lindquist, S. (2002). Neurotoxicity and neurodegeneration when PrP accumulates in the cytosol. Science298:1781–1785. [DOI] [PubMed] [Google Scholar]
  29. Manson, J. C., Clarke, A. R., Hooper, M. L., Aitchison, L., McConnell, I., and Hope, J. (1994). 129/Ola mice carrying a null mutation in PrP that abolishes mRNA production are developmentally normal. Mol. Neurobiol.8:121–127. [DOI] [PubMed] [Google Scholar]
  30. Matsumoto, A., Naito, M., Itakura, H., Ikemoto, S., Asaoka, H., Hayakawa, I., Kanamori, H., Aburatani, H., Takaku, F., Suzuki, H., Kobari, Y., Miyai, T., Takahashi, K., Cohen, E. H., Wydro, R., Housman, D. E., and Kodama, T. (1990). Human macrophage scavenger receptors: Primary structure, expression, and localization in atherosclerotic lesions. Proc. Natl. Acad. Sci. U.S.A.87:9133–9137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Mbikay, M., Seidah, N. G., and Chrétien, M. (2001). Neuroendocrine secretory protein 7B2: Structure, expression and functions. Biochem. J.357:329–342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Millward, T. A., Zolnierowicz, S., and Hemmings, B. A. (1999). Regulation of protein kinase cascades by protein phosphatase 2A. Trends Biochem. Sci.24:186–191. [DOI] [PubMed] [Google Scholar]
  33. Mironov, A., Jr., Latawiec, D., Wille, H., Bouzamondo-Bernstein, E., Legname, G., Williamson, R. A., Burton, D., DeArmond, S. J., Prusiner, S. B., and Peters, P. J. (2003). Cytosolic prion protein in neurons. J. Neurosci.23:7183–7193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Mitchell, S. A., Brown, E. C., Coldwell, M. J., Jackson, R. J., and Willis, A. E. (2001). Protein factor requirements of the Apaf-1 internal ribosome entry segment: Roles of polypyrimidine tract binding protein and upstream of N-ras. Mol. Cell. Biol.21:3364–3374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Moore, R. C., Lee, I. Y., Silverman, G. L., Harrison, P. M., Strome, R., Heinrich, C., Karunaratne, A., Pasternak, S. H., Chishti, M. A., Liang, Y., Mastrangelo, P., Wang, K., Smit, A. F. A., Katamine, S., Carlson, G. A., Cohen, F. E., Prusiner, S. B., Melton, D. W., Tremblay, P., Hood, L. E., and Westaway, D. (1999). Ataxia in prion protein (PrP)-deficient mice is associated with upregulation of the novel PrP-like protein doppel. J. Mol. Biol.292:797–817. [DOI] [PubMed] [Google Scholar]
  36. Moore, R. C., Mastrangelo, P., Bouzamondo, E., Heinrich, C., Legname, G., Prusiner, S. B., Hood, L., Westaway, D., DeArmond, S. J., and Tremblay, P. (2001). Doppl-induced cerebellar degeneration in transgenic mice. Proc. Natl. Acad. Sci. U.S.A.98:15288–15293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Nakashima, K., Yanagisawa, M., Arakawa, H., Kimura, N., Hisatsune, T., Kawabata, M., Miyazono, K., and Taga, T. (1999). Synergistic signaling in fetal brain by STAT3-smad1 complex bridged by p300. Science284:479–482. [DOI] [PubMed] [Google Scholar]
  38. Okamoto, T., Schlegel, A., Scherer, P. E., and Lisanti, M. P. (1998). Caveolins, a family of scaffolding proteins for organizing ‘‘preassembled signaling complexes’’ at the plasma membrane. J. Biol. Chem.273:5419–5422. [DOI] [PubMed] [Google Scholar]
  39. Paitel, E., da Costa, C. A., Vilette, D., Grass, J., and Checler, F. (2002). Overexpression of PrPC triggers casapase 3 activation: Potentiation by proteasome inhibitors and blockade by anti-PrP antibodies. J. Neurochem.83:1208–1214. [DOI] [PubMed] [Google Scholar]
  40. Perry, V. H., Cunningham, C., and Boche, D. (2002). Atypical inflammation in the central nervous system in prion disease. Curr. Opin. Neurol.15:349–354. [DOI] [PubMed] [Google Scholar]
  41. Pfaff, S. L., Mendelsohn, M., Stewart, C. L., Edlund, T., and Jessel, T. M. (1996). Requirement for ILM homeobox gene Isl1 in motor neuron generation reveals a motor neuron-dependent step in interneuron differentiation. Cell84:309–320. [DOI] [PubMed] [Google Scholar]
  42. Prusiner, S. B. (1998). Prions. Proc. Natl. Acad. Sci. U.S.A.95:13363–13383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Rachidi, W., Vilette, D., Guiraud, P., Arlotto, M., Riondel, J., Laude, H., Lehmann, S., and Favier, A. (2003). Expression of prion protein increases cellular copper binding and antioxidant enzyme activities but not copper delivery. J. Biol. Chem.278:9064–9072. [DOI] [PubMed] [Google Scholar]
  44. Rossi, D., Cozzio, A., Flechsig, E., Klein, M. A., Rülicke, T., Aguzzi, A., and Weissmann, C. (2001). Onset of ataxia and Purkinje cell loss in PrP null mice inversely correlated with Dpl level in brain. EMBO J.20:694–702. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Sakaguchi, S., Katamine, S., Nishida, N., Moriuchi, R., Shigematsu, K., Sugimoto, T., Nakatani, A., Kataoka, Y., Houtani, T., Shirabe, S., Okada, H., Hasegawa, S., Miyamoto, T., and Noda, T. (1996). Loss of cerebellar Purkinje cells in aged mice homozygous for a disrupted PrP gene. Nature380:528–531. [DOI] [PubMed] [Google Scholar]
  46. Satoh, J.-I., and Kuroda, Y. (2000). Differential gene expression between human neurons and neuronal progenitor cells in culture: An analysis of arrayed cDNA clones in NTera2 human embryonal carcinoma cell line as a model system. J. Neurosci. Methods94:155–164. [DOI] [PubMed] [Google Scholar]
  47. Satoh, J.-I., Kuroda, Y., and Katamine, S. (2000). Gene expression profile in prion protein-deficient fibroblasts in culture. Am. J. Pathol.157:59–68. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Satoh, J.-I., Yukitake, M., Kurohara, K., Nishida, N., Katamine, S., Miyamoto, T., and Kuroda, Y. (1998). Cultured skin fibroblasts isolated from mice devoid of the prion protein gene express major heat shock proteins in response to heat stress. Exp. Neurol.151:105–115. [DOI] [PubMed] [Google Scholar]
  49. Schmidt, K., Kins, S., Schild, A., Nitsch, R. M., Hemmings, B. A., and Götz, J. (2002). Diversity, developmental regulation and distribution of murine PR55/B subunits of protein phosphatase 2A. Eur. J. Neurosci.16:2039–2048. [DOI] [PubMed] [Google Scholar]
  50. Schmitt-Ulms, G., Legname, G., Baldwin, M. A., Ball, H. L., Bradon, N., Bosque, P. J., Crossin, K. L., Edelman, G. M., DeArmond, S. J., Cohen, F. E., and Prusiner, S. B. (2001). Binding of neural cell adhesion molecules (N-CAMs) to the cellular prion protein. J. Mol. Biol.314:1209–1225. [DOI] [PubMed] [Google Scholar]
  51. Shmakov, A. N., McLennan, N. F., McBride, P., Farqyhar, C. F., Bode, J., Pennison, K. A., and Ghosh, S. (2000). Cellular prion protein is expressed in the human enteric nervous system. Nature Med.6:840–841. [DOI] [PubMed] [Google Scholar]
  52. Simmons, A. D., Püschel, A. W., McPherson, J. D., Overhauser, J., and Lovett, M. (1998). Molecular cloning and mapping of human semaphorin F from the Cri-du-chat candidate interval. Biochem. Biophys. Res. Commun.242:685–691. [DOI] [PubMed] [Google Scholar]
  53. Sledz, C. A., Holko, M., de Veer, M. J., Silverman, R. H., and Williams, B. R. G. (2003). Activation of the interferon system by short-interfering RNAs. Nature Cell Biol.5:834–839. [DOI] [PubMed] [Google Scholar]
  54. Sontag, E., Nunbhakdi-Craig, V., Lee, G., Brandt, R., Kamibayashi, C., Kuret, J., White, C. L., III, Mumby, M. C., and Bloom, G. S. (1999). Molecular interactions among protein phosphatase 2A, tau, and microtubules. J. Biol. Chem.274:25490–25498. [DOI] [PubMed] [Google Scholar]
  55. Tezak, Z., Prandini, P., Boscaro, M., Marin, A., Devaney, J., Marino, M., Fanin, M., Trevisan, C. P., Park, J., Tyson, W., Finkel, R., Garcia, C., Angelini, C., Hoffman, E. P., and Pegoraro, E. (2003). Clinical and molecular study in congenital muscular dystrophy with partial laminin α2 (LAMA2) deficiency. Hum. Mut.21:103–111. [DOI] [PubMed] [Google Scholar]
  56. Tobler, I., Gaus, S. E., Deboer, T., Achermann, P., Fischer, M., Rülicke, T., Moser, M., Oesch, B., McBride, P. A., and Manson, J. C. (1996). Altered circadian activity rhythms and sleep in mice devoid of prion protein. Nature380:639–642. [DOI] [PubMed] [Google Scholar]
  57. Vey, M., Pilkuhn, S., Wille, H., Nixon, R., DeArmond, S. J., Smart, E. J., Anderson, R. G. W., Taraboulos, A., and Prusiner, S. B. (1996). Subcellular colocalization of the cellular and scrapie prion proteins in caveolae-like membranous domains. Proc. Natl. Acad. Sci. U.S.A.93:14945–14949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Wang, X.-Y., Smith, D. I., Liu, W., and James, C. D. (1998). GBAS, a novel gene encoding a protein with tyrosine phosphorylation sites and a transmembrane domain is co-purified with EGFR. Genomics49:448–451. [DOI] [PubMed] [Google Scholar]
  59. Westaway, D., DeArmond, S. J., Cayetano-Canlas, J., Groth, D., Foster, D., Yang, S. L., Torchia, M., Carlson, G. A., and Prusiner, S. B. (1994). Degeneration of skeletal muscle, peripheral nerves, and the central nervous system in transgenic mice overexpressing wild-type prion proteins. Cell76:117–129. [DOI] [PubMed] [Google Scholar]
  60. Yamaguchi, T., Murata, Y., Fujiyoshi, Y., and Doi, T. (2003). Regulated interaction of endothelin B receptor with caveolin-1. Eur. J. Biochem.270:1816–1827. [DOI] [PubMed] [Google Scholar]

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