The N-Terminal Polybasic Region of Prion Protein Is Crucial in Prion Pathogenesis Independently of the Octapeptide Repeat Region

  • Nandita Rani Das
  • Hironori Miyata
  • Hideyuki Hara
  • Junji Chida
  • Keiji Uchiyama
  • Kentaro Masujin
  • Hitomi Watanabe
  • Gen Kondoh
  • Suehiro SakaguchiEmail author


Conformational conversion of the cellular isoform of prion protein, designated PrPC, into the abnormally folded, amyloidogenic isoform, PrPSc, is an essential pathogenic event in prion diseases. However, the exact conversion mechanism remains largely unknown. Lines of evidence indicate that the N-terminal domain, which includes the N-terminal, positively charged polybasic region and the octapeptide repeat (OR) region, is important for PrPC to convert into PrPSc after infection with prions. To further gain insights into the role of the polybasic region and the OR region in prion pathogenesis, we generated two different transgenic mice, designated Tg(PrP3K3A)/Prnp0/0 and Tg(PrP3K3A∆OR)/Prnp0/0 mice, which express PrPC with lysine residues at codons 23, 24, and 27 in the polybasic region mutated with or without a deletion of the OR region on the Prnp0/0 background, respectively, and intracerebrally inoculated them with RML and 22L prions. We show that Tg(PrP3K3A)/Prnp0/0 mice were highly resistant to the prions, indicating that lysine residues at 23, 24, and 27 could be important for the polybasic region to support prion infection. Tg(PrP3K3A∆OR)/Prnp0/0 mice also had reduced susceptibility to RML and 22L prions equivalent to Tg(PrP3K3A)/Prnp0/0 mice. The pre-OR region, including the polybasic region, of PrP3K3A∆OR, but not PrP3K3A, was unusually converted to a protease-resistant structure during conversion to PrPSc3K3A∆OR. These results suggest that, while the OR region could affect the conformation of the polybasic region during conversion of PrPC into PrPSc, the polybasic region could play a crucial role in prion pathogenesis independently of the OR region.


Prion Prion protein Polybasic region Octapeptide repeat Transgenic mice Neurodegeneration 



We thank Dr. Stanley B. Prusiner for providing Zrch I Prnp0/0 mice.

Authors’ Contributions

SS designed the experiments. NRD and HM performed prion infection. HH performed pathological experiments. NRD, HH, JC, and KU performed biochemical experiments. KM performed an epitope mapping. HM, HW, and GK produced Tg mice. SS wrote the paper. All authors revised and approved the final manuscript.

Funding Information

This work was supported in part by JSPS KAKENHI 26293212 and 19H03548, MEXT KAKENHI 15H01560, and 17H05701 to SS.

Compliance with Ethical Standards

All experiments were performed in accordance with The Guiding Principle for Animal Care and Experimentation of the University of Occupational and Environmental Health and Tokushima University and with Japanese Law for Animal Welfare and Care. The Ethics Committees of Animal Care and Experimentation of the University of Occupational and Environmental Health and Tokushima University approved this study (approval number AE08-013, T28-100). All efforts were made to minimize animal suffering and to reduce the number of animals used.

Conflict of Interest

The authors declare that they have no conflicts of interest.


  1. 1.
    Stahl N, Borchelt DR, Hsiao K, Prusiner SB (1987) Scrapie prion protein contains a phosphatidylinositol glycolipid. Cell 51(2):229–240CrossRefGoogle Scholar
  2. 2.
    Oesch B, Westaway D, Walchli M, McKinley MP, Kent SB, Aebersold R, Barry RA, Tempst P et al (1985) A cellular gene encodes scrapie PrP 27-30 protein. Cell 40(4):735–746CrossRefGoogle Scholar
  3. 3.
    Prusiner SB (1998) Prions. Proc Natl Acad Sci U S A 95(23):13363–13383CrossRefGoogle Scholar
  4. 4.
    Weissmann C, Enari M, Klohn PC, Rossi D, Flechsig E (2002) Molecular biology of prions. Acta Neurobiol Exp 62(3):153–166Google Scholar
  5. 5.
    Bueler H, Aguzzi A, Sailer A, Greiner RA, Autenried P, Aguet M, Weissmann C (1993) Mice devoid of PrP are resistant to scrapie. Cell 73(7):1339–1347CrossRefGoogle Scholar
  6. 6.
    Prusiner SB, Groth D, Serban A, Koehler R, Foster D, Torchia M, Burton D, Yang SL et al (1993) Ablation of the prion protein (PrP) gene in mice prevents scrapie and facilitates production of anti-PrP antibodies. Proc Natl Acad Sci U S A 90(22):10608–10612CrossRefGoogle Scholar
  7. 7.
    Manson JC, Clarke AR, McBride PA, McConnell I, Hope J (1994) PrP gene dosage determines the timing but not the final intensity or distribution of lesions in scrapie pathology. Neurodegener 3(4):331–340Google Scholar
  8. 8.
    Sakaguchi S, Katamine S, Shigematsu K, Nakatani A, Moriuchi R, Nishida N, Kurokawa K, Nakaoke R et al (1995) Accumulation of proteinase K-resistant prion protein (PrP) is restricted by the expression level of normal PrP in mice inoculated with a mouse-adapted strain of the Creutzfeldt-Jakob disease agent. J Virol 69(12):7586–7592PubMedPubMedCentralGoogle Scholar
  9. 9.
    Turnbaugh JA, Unterberger U, Saa P, Massignan T, Fluharty BR, Bowman FP, Miller MB, Supattapone S et al (2012) The N-terminal, polybasic region of PrP(C) dictates the efficiency of prion propagation by binding to PrP(Sc). J Neurosci 32(26):8817–8830. CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Khalife M, Reine F, Paquet-Fifield S, Castille J, Herzog L, Vilotte M, Moudjou M, Moazami-Goudarzi K et al (2016) Mutated but not deleted ovine PrP(C) N-terminal polybasic region strongly interferes with prion propagation in transgenic mice. J Virol 90(3):1638–1646. CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Yamaguchi Y, Miyata H, Uchiyama K, Ootsuyama A, Inubushi S, Mori T, Muramatsu N, Katamine S et al (2012) Biological and biochemical characterization of mice expressing prion protein devoid of the octapeptide repeat region after infection with prions. PLoS One 7(8):e43540. CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Ishibashi D, Yamanaka H, Yamaguchi N, Yoshikawa D, Nakamura R, Okimura N, Yamaguchi Y, Shigematsu K et al (2007) Immunization with recombinant bovine but not mouse prion protein delays the onset of disease in mice inoculated with a mouse-adapted prion. Vaccine 25(6):985–992CrossRefGoogle Scholar
  13. 13.
    Yoshikawa D, Yamaguchi N, Ishibashi D, Yamanaka H, Okimura N, Yamaguchi Y, Mori T, Miyata H et al (2008) Dominant-negative effects of the N-terminal half of prion protein on neurotoxicity of prion protein-like protein/doppel in mice. J Biol Chem 283(35):24202–24211CrossRefGoogle Scholar
  14. 14.
    Brinster RL, Chen HY, Trumbauer ME, Yagle MK, Palmiter RD (1985) Factors affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs. Proc Natl Acad Sci U S A 82(13):4438–4442CrossRefGoogle Scholar
  15. 15.
    Wilmut I, Hooper ML, Simons JP (1991) Genetic manipulation of mammals and its application in reproductive biology. J Reprod Fertil 92(2):245–279CrossRefGoogle Scholar
  16. 16.
    Scott MR, Kohler R, Foster D, Prusiner SB (1992) Chimeric prion protein expression in cultured cells and transgenic mice. Protein Sci 1(8):986–997CrossRefGoogle Scholar
  17. 17.
    Feraudet C, Morel N, Simon S, Volland H, Frobert Y, Creminon C, Vilette D, Lehmann S et al (2005) Screening of 145 anti-PrP monoclonal antibodies for their capacity to inhibit PrPSc replication in infected cells. J Biol Chem 280(12):11247–11258CrossRefGoogle Scholar
  18. 18.
    Scott M, Foster D, Mirenda C, Serban D, Coufal F, Walchli M, Torchia M, Groth D et al (1989) Transgenic mice expressing hamster prion protein produce species-specific scrapie infectivity and amyloid plaques. Cell 59(5):847–857CrossRefGoogle Scholar
  19. 19.
    Das NR, Miyata H, Hara H, Uchiyama K, Chida J, Yano M, Watanabe H, Kondoh G et al (2017) Effects of prion protein devoid of the N-terminal residues 25-50 on prion pathogenesis in mice. Arch Virol 162(7):1867–1876. CrossRefPubMedGoogle Scholar
  20. 20.
    Pan T, Wong BS, Liu T, Li R, Petersen RB, Sy MS (2002) Cell-surface prion protein interacts with glycosaminoglycans. Biochem J 368(Pt 1):81–90CrossRefGoogle Scholar
  21. 21.
    Warner RG, Hundt C, Weiss S, Turnbull JE (2002) Identification of the heparan sulfate binding sites in the cellular prion protein. J Biol Chem 277(21):18421–18430CrossRefGoogle Scholar
  22. 22.
    Taubner LM, Bienkiewicz EA, Copie V, Caughey B (2010) Structure of the flexible amino-terminal domain of prion protein bound to a sulfated glycan. J Mol Biol 395(3):475–490. CrossRefPubMedGoogle Scholar
  23. 23.
    Taylor DR, Watt NT, Perera WS, Hooper NM (2005) Assigning functions to distinct regions of the N-terminus of the prion protein that are involved in its copper-stimulated, clathrin-dependent endocytosis. J Cell Sci 118(Pt 21):5141–5153CrossRefGoogle Scholar
  24. 24.
    Walmsley AR, Zeng F, Hooper NM (2003) The N-terminal region of the prion protein ectodomain contains a lipid raft targeting determinant. J Biol Chem 278(39):37241–37248CrossRefGoogle Scholar
  25. 25.
    Caughey B, Raymond GJ, Ernst D, Race RE (1991) N-terminal truncation of the scrapie-associated form of PrP by lysosomal protease(s): implications regarding the site of conversion of PrP to the protease-resistant state. J Virol 65(12):6597–6603PubMedPubMedCentralGoogle Scholar
  26. 26.
    Borchelt DR, Taraboulos A, Prusiner SB (1992) Evidence for synthesis of scrapie prion proteins in the endocytic pathway. J Biol Chem 267(23):16188–16199PubMedGoogle Scholar
  27. 27.
    Wu B, McDonald AJ, Markham K, Rich CB, McHugh KP, Tatzelt J, Colby DW, Millhauser GL et al (2017) The N-terminus of the prion protein is a toxic effector regulated by the C-terminus. Elife:6.

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Division of Molecular Neurobiology, The Institute for Enzyme Research (KOSOKEN)Tokushima UniversityTokushimaJapan
  2. 2.Animal Research Center, School of MedicineUniversity of Occupational and Environmental HealthKitakyushuJapan
  3. 3.Influenza and Prion Disease Research CenterNational Institute of Animal Health (NIAH), National Agriculture and Food Research Organization (NARO)IbarakiJapan
  4. 4.Laboratory of Integrative Biological Science, Institute for Frontier Life and Medical SciencesKyoto UniversityKyotoJapan

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