Skip to main content
Log in

Cloning of the bovine prion-like Shadoo (SPRN) gene by comparative analysis of the predicted genomic locus

  • Published:
Mammalian Genome Aims and scope Submit manuscript

Abstract

SPRN is a new prion-like gene coding for Sho, a protein with significant similarity to PrP. SPRN was initially described in zebrafish; however, the strong evolutionary conservation led to the hypothesis that SPRN might be the ancestral prion-like gene. We mapped SPRN in Bos taurus by comparative analysis of the locus and of the predicted flanking genes. BACs, spanning the whole SPRN genomic locus, were assigned to BTA26q23 by radiation hybrid mapping and fluorescent in situ hybridization (FISH). Sequencing of five genes flanking SPRN, namely, ECHS1, PAOX, MTG1, SPRN, and CYP2E1, high-resolution FISH on mechanically stretched chromosomes, and combed BAC DNA allowed us to establish their order and reciprocal orientation. The results confirmed that BTA26q23 corresponds to HSA10q24.3–26.3, which is the site where the human SPRN is located. The gene order in Bos taurus is the same as in man, cen-ECHS1-PAOX-MTG1-SPRN-CYP2E1-tel, but PAOX has a different orientation in the two species. SPRN has the typical two-exon PRNP arrangement, with the CDS fully contained within exon 2; furthermore, it codes for a 143-amino-acid protein with 74.8% identity and 84.7% similarity with the human PRNP. RT-PCR and Northern blot analysis showed that SPRN is expressed at high levels in brain and less in testis and lung.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1.
Fig. 2.
Fig. 3.

Similar content being viewed by others

References

  • Castiglioni B, Comincini S, Drisaldi B, Motta T, Ferretti L (1998) Comparative mapping of the prion gene (PRNP) locus in cattle, sheep and human with PCR-generated probes. Mamm Genome 9, 853–855

    Article  PubMed  CAS  Google Scholar 

  • Choi SH, Kim IC, Kim DS, Kim DW, Chae SH, et al. (2006) Comparative genomic organization of the human and bovine PRNP locus. Genomics 87, 598–607

    Article  PubMed  CAS  Google Scholar 

  • Comincini S, Foti MG, Tranulis MA, Hills D, Di Guardo G, et al. (2001) Genomic organization, comparative analysis, and genetic polymorphisms of the bovine and ovine prion Doppel genes (PRND). Mamm Genome 12, 729–733

    Article  PubMed  CAS  Google Scholar 

  • de Givry S, Bouchez M, Chabrier P, Milan D, Schiex T (2005) CARHTA GENE: multipopulation integrated genetic and radiation hybrid mapping. Bioinformatics 21, 1703–1704

    Article  PubMed  CAS  Google Scholar 

  • Eggen A, Gautier M, Billaut A, Petit E, Hayes H, et al. (2001) Construction and characterization of a bovine BAC library with four genome-equivalent coverage. Genet Sel Evol 33, 543–548

    Article  PubMed  CAS  Google Scholar 

  • Gabriel JM, Oesch B, Kretzschmar H, Scott M, Prusiner SB (1992) Molecular cloning of a candidate chicken prion protein. Proc Natl Acad Sci U S A 89, 9097–9101

    Article  PubMed  CAS  Google Scholar 

  • Gautier M, Hayes H, Taourit S, Laurent P, Eggen A (2001) Syntenic assignment of sixteen genes from the long arm of human chromosome 10 to bovine chromosomes 26 and 28: refinement of the comparative map. Chromosome Res 9, 617–621

    Article  PubMed  CAS  Google Scholar 

  • Gautier M, Hayes H, Eggen A (2003) A comprehensive radiation hybrid map of bovine Chromosome 26 (BTA26): comparative chromosomal organization between HSA10q and BTA26 and BTA28. Mamm Genome 14, 711–721

    Article  PubMed  CAS  Google Scholar 

  • Haaf T, Ward DC (1994) High resolution ordering of YAC contigs using extended chromatin and chromosomes. Hum Mol Genet 3, 629–633

    PubMed  CAS  Google Scholar 

  • Henegariu O, Grober L, Haskins W, Bowers PN, State MW, et al. (2001) Rapid DNA fiber technique for size measurements of linear and circular DNA probes. Biotechniques 31, 246–250

    PubMed  CAS  Google Scholar 

  • Hills D, Comincini S, Schlaepfer J, Dolf G, Ferretti L, et al. (2001) Complete genomic sequence of the bovine prion gene (PRNP) and polymorphism in its promoter region. Anim Genet 32, 231–232

    Article  PubMed  CAS  Google Scholar 

  • Iannuzzi L, Perucatti A, Di Meo GP, Schibler L, Incarnato D, et al. (2003) Chromosomal localization of sixty autosomal loci in sheep (OVIS ARIES, 2n = 54) by fluorescence in situ hybridization and R-banding. Cytogenet Genome Res 103, 135–138

    Article  PubMed  CAS  Google Scholar 

  • Lee IY, Westaway D, Smit AF, Wang K, Seto J, et al. (1998) Complete genomic sequence and analysis of the prion protein gene region from three mammalian species. Genome Res 8, 1022–1037

    PubMed  CAS  Google Scholar 

  • Li G, Bolton DC (1997) A novel hamster prion protein mRNA contains an extra exon: increased expression in scrapie. Brain Res 751, 265–274

    Article  PubMed  CAS  Google Scholar 

  • Makrinou E, Collinge J, Antoniou M (2002) Genomic characterization of the human prion protein (PrP) gene locus. Mamm Genome 13, 696–703

    Article  PubMed  CAS  Google Scholar 

  • Moore RC, Lee IY, Silverman GL, Harrison PM, Strome R, et al. (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

    Article  PubMed  CAS  Google Scholar 

  • Oidtmann B, Simon D, Holtkamp N, Hoffmann R, Baier M (2003) Identification of cDNAs from Japanese pufferfish (Fugu rubripes) and Atlantic salmon (Salmo salar) coding for homologues to tetrapod prion proteins. FEBS Lett 538, 96–100

    Article  PubMed  CAS  Google Scholar 

  • Premzl M, Sangiorgio L, Strumbo B, Marshall Graves JA, et al. (2003) Shadoo, a new protein highly conserved from fish to mammals and with similarity to prion protein. Gene 314, 89–102

    Article  PubMed  CAS  Google Scholar 

  • Premzl M, Gready JE, Jermiin LS, Simonic T, Marshall Graves JA (2004) Evolution of vertebrate genes related to Prion and Shadoo proteins – clues from comparative genomic analysis. Mol Biol Evol 21, 2210–2231

    Article  PubMed  CAS  Google Scholar 

  • Raimondi E, Ferretti L, Young BD, Sgaramella V, De Carli L (1991) The origin of a morphologically unidentifiable human supernumerary minichromosome traced via sorting molecular cloning and in situ hybridization. J Med Genet 28, 92–96

    Article  PubMed  CAS  Google Scholar 

  • Rivera–Milla E, Stuermer CA, Malaga–Trillo E (2003) An evolutionary basis for scrapie disease: identification of a fish prion mRNA. Trends Genet 19, 72–75

    Article  PubMed  CAS  Google Scholar 

  • Rivera–Milla E, Oidtmann B, Panagiotidis CH, Baier M, Sklaviadis T, et al. (2006) Disparate evolution of prion protein domains and the distinct origin of Doppel- and prion-related loci revealed by fish-to-mammal comparisons. FASEB J 20, 317–319

    PubMed  CAS  Google Scholar 

  • Simonic T, Duga S, Strumbo B, Asselta R, Ceciliani F, et al. (2000) cDNA cloning of turtle prion protein. FEBS Lett 469, 33–38

    Article  PubMed  CAS  Google Scholar 

  • Strumbo B, Ronchi S, Bolis LC, Simonic T (2001) Molecular cloning of the cDNA coding for Xenopus laevis prion protein. FEBS Lett 508, 170–174

    Article  PubMed  CAS  Google Scholar 

  • Suzuki T, Kurokawa T, Hashimoto H, Sugiyama M (2002) cDNA sequence and tissue expression of Fugu rubripes prion protein-like: a candidate for the teleost orthologue of tetrapod PrPs. Biochem Biophys Res Commun 294, 912–917

    Article  PubMed  CAS  Google Scholar 

  • Uboldi C, Del Vecchio I, Foti MG, Azzalin A, Paulis M, et al. (2005) Prion-like Doppel gene (PRND) in the goat: genomic structure, cDNA, and polymorphisms. Mamm Genome 16, 963–971

    Article  PubMed  CAS  Google Scholar 

  • Williams JL, Eggen A, Ferretti L, Farr CJ, Gautier M, et al. (2002) A bovine whole-genome radiation hybrid panel and outline map. Mamm Genome 13, 469–474

    Article  PubMed  CAS  Google Scholar 

  • Windl O, Dempster M, Estibeiro P, Lathe R (1995) A candidate marsupial PrP gene reveals two domains conserved in mammalian PrP proteins. Gene 159, 181–186

    Article  PubMed  CAS  Google Scholar 

  • Wopfner F, Weidenhofer G, Schneider R, von Brunn A, Gilch S, et al. (1999) Analysis of 27 mammalian and 9 avian PrPs reveals high conservation of flexible regions of the prion protein. J Mol Biol 289, 1163–1178

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the Italian MIUR, Progetti di Ricerca di Rilevante Interesse Nazionale (2005), and from the Istituto Superiore di Sanità (ISS) “UO9 – Fattori genetici, patogenetici e biochimici responsabili della sensibilità/resistenza alle EST.”

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Luca Ferretti.

Electronic Supplementary Material

Rights and permissions

Reprints and permissions

About this article

Cite this article

Uboldi, C., Paulis, M., Guidi, E. et al. Cloning of the bovine prion-like Shadoo (SPRN) gene by comparative analysis of the predicted genomic locus. Mamm Genome 17, 1130–1139 (2006). https://doi.org/10.1007/s00335-006-0078-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00335-006-0078-7

Keywords

Navigation