Mammalian Genome

, Volume 4, Issue 11, pp 662–669

A physical map of the human APP gene in YACs

  • Karen Rooke
  • Chris Talbot
  • Louise James
  • Rakesh Anand
  • John A. Hardy
  • Alison M. Goate
Original Contributions

Abstract

Several point mutations within exons 16 and 17 of the amyloid precursor protein (APP) gene have been reported that are associated with Alzheimer's disease in a small number of familial cases. To determine the size of the APP gene and the organization of the exons within human genomic DNA, we have characterized 11 Yeast Artificial Chromosome (YAC), recombinants containing human APP gene sequences. The smallest YAC insert was 125 kb, and the largest was 1.4 Mb. The YACs were screened by polymerase chain reaction amplification of APP exons to determine which of the 18 exons coding for APP770 were present. Four of the YACs (D110G1, D110G6, D110E9, and B142F9) contain all 18 exons and at least part of the promoter. Construction of an overlapping map of the gene with all of the YACs demonstrated that 3 of the 11 YACs were chimeric. The orientation and position of the coding sequence on the map was determined by probing digests of the YAC DNA with exon PCR products and the vector arms. The coding region of the APP gene spans approximately 400 kb of genomic DNA.

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References

  1. Anand, R., Villasante, A., Tyler-Smith, C. (1989). Construction of yeast artificial chromosome libraries with large inserts using fractionation by pulsed-field gel electrophoresis. Nucleic Acids Res. 17, 3425–3433.Google Scholar
  2. Anand, R., Riley, J., Smith, J., Markham, A. (1990). A 3.5 genome equivalent multi-access YAC library: construction, characterisation, screening and storage. Nucleic Acids Res. 18, 1951–1956.Google Scholar
  3. Burke, D., Carle, G., Olson, M. (1987). Cloning of large segments of exogenous DNA into yeast by means of artificial chromosomes. Science 236, 806–812.Google Scholar
  4. Chartier-Harlin, M.C., Crawford, F., Houlden, H., Warren, A., Hughes, D., Fidani, L., Goate, A., Rossor, M., Roques, P., Hardy, J., Mullan, M. (1991). Early onset Alzheimer's disease caused by mutations at codon 717 of the β-amyloid precursor protein gene. Nature 353, 844–846.Google Scholar
  5. Chu, G., Vollrath, D., Davis, R. (1986). Separation of large DNA molecules by contour-clamped homogeneous electric fields. Science 234, 1582–1585.Google Scholar
  6. Church, G., Gilbert, W. (1984). Genomic sequencing. Proc. Natl. Acad. Sci. USA 81, 1991–1995.Google Scholar
  7. De Sauvage, F., Octave, J.N. (1989). A novel mRNA of the A4 amyloid precursor gene coding for a possibly secreted protein. Science 245, 651–653.Google Scholar
  8. Feinberg, A., Vogelstein, B. (1983). A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 132, 6–13.Google Scholar
  9. Fidani, L., Rooke, K., Chartier-Harlin, M.C., Hughes, D., Tanzi, R., Mullan, M., Roques, P., Rossor, M., Hardy, J., Goate, A.M. (1992). Screening for mutations in the open reading frame and promoter of the β-amyloid precursor protein gene in familial Alzheimer's disease: identification of a further family with APP717 Val-> Ile. Hum. Mol. Genet. 1, 165–168.Google Scholar
  10. Goate, A., Chartier-Harlin, M.C., Mullan, M., Brown, J., Crawford, F., Fidani, L., Giuffra, L., Haynes, A., Irving, N., James, L., Mant, R., Newton, P., Rooke, K., Roques, P., Talbot, C., Pericak-Vance, M., Roses, A., Williamson, R., Rossor, M., Owen, M., Hardy, J. (1991). Segregation of a missense mutation in the amyloid precursor protein gene with familial Alzheimer's disease. Nature 349, 704–706.Google Scholar
  11. Goate, A.M., Haynes, A., Owen, M.J., Farrall, M., James, L.A., Lai, L.Y., Mullan, M.J., Roques, P., Rossor, M.N., Williamson, R., Hardy, J. (1989). Predisposing locus for Alzheimer's disease on chromosome 21. Lancet 1, 352–355.Google Scholar
  12. Golde, T.E., Estus, S., Usiak, M., Younkin, L.H., Younkin, S.G. (1990). Expression of beta amyloid protein precursor mRNAs: recognition of a novel alternatively spliced form and quantitation in Alzheimer's disease using PCR. Neuron 4, 253–267.Google Scholar
  13. Goldgaber, D., Lerman, M., McBride, W.O., Saffiotti, U., Gajdusek, D.C. (1987). Isolation and chromosomal localization of human brain cDNA clones coding for the precursor of the amyloid of brain in Alzheimer's disease, Down's syndrome and aging. Science 235, 877–880.Google Scholar
  14. Herrmann, B., Barlow, D., Lehrach, H. (1987). A large inverted duplication allows homologous recombination between chromosomes heterozygous for the proximal t complex inversion. Cell 48, 813–825.Google Scholar
  15. Kang, J., Lemaire, H., Unterbeck, A., Salbaum, J.M., Masters, C.L., Grzeschik, K.H., Multhaup, G., Beyreuther, K., MullerHill, B. (1987). The precursor of Alzheimer's disease amyloid A4 protein resembles a cell-surface receptor. Nature 325, 733–736.Google Scholar
  16. Karlinsky, H., Vaula, G., Haines, J.L., Ridgley, J., Bergeron, C., Mortilla, M., Tupler, R.G., Percy, M.E., Robitaille, Y., Noldy, N.E., Yip, T.C.K., Tanzi, R.E., Gusella, J.F., Becker, R., Berg, J.M., Crapper-McLachlan, D.R., St. George-Hyslop, P.H. (1992). Molecular and prospective phenotypic characterization of a pedigree with familial Alzheimer's disease and a missense mutation in codon 717 of the β-amyloid precursor protein gene. Neurology 42, 1445–1453.Google Scholar
  17. Kitaguchi, N., Takahashi, Y., Tokushima, Y., Shiojiri, S., Ito, H. (1988). Novel precursor of Alzheimer's disease amyloid protein shows protease inhibitory activity. Nature 331, 530–532.Google Scholar
  18. Levy, E., Carman, M., Fernandez-Madrid, I.J., Power, M.D., Lieberburg, I., can Duinen, S.G., Bots, G.T., Luyendijk, W., Frangione, B. (1990). Mutation of the Alzheimer's disease amyloid gene in hereditary cerebral hemorrhage, Dutch type. Science 248, 1124–1126.Google Scholar
  19. Mann, D.M. (1988). Alzheimer's disease and Down's syndrome. Histopathology 13, 125–137.Google Scholar
  20. Mullan, M., Crawford, F., Axelman, K., Houlden, H., Lilius, L., Winblad, B., Lannfelt, L. (1992). A new mutation in APP demonstrates that pathogenic mutations for probable Alzheimer's disease frame the β-amyloid sequence. Nature Genet. 1, 345–347.Google Scholar
  21. Murrell, J., Farlow, M., Ghetti, B., Benson, M. (1991). A mutation in the amyloid precursor protein associated with hereditary Alzheimer's disease. Science 254, 97–99.Google Scholar
  22. Naruse, S., Igarashi, S., Kobayashi, H., Aoki, K., Inuzuka, T., Kaneko, K., Shimizu, T., Iihara, K., Kojima, T., Miyatake, T., Tsuji, S. (1991). Mis-sense mutation Val-> Ile in exon 17 of amyloid precursor protein gene in Japanese familial Alzheimer's disease. Lancet 337, 978–979.Google Scholar
  23. Orr-Weaver, T., Szostak, J., Rothstein, R. (1983). Genetic applications of yeast transformation with linear and gapped plasmids. Methods Enzymol. 101, 228–245.Google Scholar
  24. Riley, J., Ogilvie, D., Anand, R. (1992). Construction, characterization and screening YAC libraries. In Techniques for the Analysis of Complex Genomes, R. Anand, ed. (New York: Academic Press), pp. 59–79.Google Scholar
  25. Rumble, B., Retallack, R., Hilbich, C., Simms, G., Multhaup, G., Martins, R., Hockey, A., Montgomery, P., Beyreuther, K., Masters, C.L. (1989). Amyloid A4 protein and its precursor in Down's syndrome and Alzheimer's disease. N. Engl. J. Med. 320, 1446–1452.Google Scholar
  26. Salbaum, J.M., Weidemann, A., Lemaire, H.G., Masters, C.L., Beyreuther, K. (1988). The promoter of Alzheimer's disease amyloid A4 precursor gene. EMBO J. 7, 2807–2813.Google Scholar
  27. Schellenberg, G.D., Anderson, L., O'Dahl, S., Wijsman, E., Sadovnick, A.D., Ball, M.J., Larson, E.B., Kukull, W.A., Martin, G.M., Roses, A.D., Bird, T.D. (1991). APP717, APP693, and PRIP gene mutations are rare in Alzheimer's disease. Am. J. Hum. Genet. 49, 511–517.Google Scholar
  28. Scherer, S., Davis, r. (1979). Replacement of chromosome segments with altered DNA sequences constructed in vitro. Proc. Natl. Acad. Sci. USA 76, 4951–4955.Google Scholar
  29. Sorbi, S., Nacmias, B., Forleo, P., Piacentini, S., Amaducci, L., Provinciali, L. (1993). APP717 and Alzheimer's disease in Italy. Nature Genet, 4, 10.Google Scholar
  30. St. George-Hyslop, P.H., Haines, J., Farrer, L.A., Polinsky, R., Van Broeckhoven, C., Goate, A., McLachlan, D.R., Orr, H., Bruni, A.C., Sorbi, S., Rainero, I., Foncin, J., Pollen, D., Cantu, J., Tupler, R., Voskresenskaya, N., Mayeux, R., Growdon, J., Fried, V., Myers, R., Nee, L., Backhovens, H., Martin, J-J., Rossor, M., Owen, M., Mullan, M., Percy, M., Karlinsky, H., Rich, S., Heston, L., Montesi, M., Mortilla, M., Nacmias, N., Gusella, J., Hardy, J. (1990). Genetic linkage studies suggest that Alzheimer's disease is not a single homogeneous disorder. FAD Collaboative Study Group. Nature 347, 194–197.Google Scholar
  31. Tanzi, R., Vaula, G., Romano, D., Mortilla, M., Huang, T., Tupler, R., Wasco, W., Hyman, B., Haines, J., Jenkins, B., Kalaitsidaki, M., Warren, A., McGinnis, M., Antonorakis, S., Karlinsky, H., Percy, M., Connor, L., Growdon, J., Crapper-Mclachlan, D., Gusella, J., St George-Hyslop, P. (1992). Assessment of amyloid beta-protein precursor gene mutations in a large set of familial and sporadic Alzheimer disease cases. Am. J. Hum. Genet. 51, 273–282.Google Scholar
  32. Tanzi, R.E., Gusella, J., Watkins, P.C., Bruns, G.A., St George-Hyslop, P., Van Keuren, M.L., Patterson, D., Pagan, S., Kurnit, D.M., Neve, R.L. (1987). Amyloid beta protein gene: cDNA, mRNA distribution, and genetic linkage near the Alzheimer locus. Science 235, 880–884.Google Scholar
  33. Tanzi, R.E., McClatchey, A.I., Lamberti, E.D., Villa Komaroff, L., Gusella, J.F., Neve, R.L. (1988). Protease inhibitor domain encoded by an amyloid precursor mRNA associated with Alzheimer's disease. Nature 331, 528–530.Google Scholar
  34. Tanzi, R.E., McClatchey, A.I., Lamberti, E.D., Villa Komaroff, L., Gusella, J.F., Neve, R.L. (1992). Protease inhibitor domain encoded by an amyloid precursor mRNA associated with Alzheimer's disease. Nature 331, 528–530.Google Scholar
  35. Tassoni, F., Cheng, S., Gardiner, K. (1992). Analysis of chromosome 21 yeast artificial chromosome (YAC) clones. Am. J. Hum. Genet. 51, 1251–1264.Google Scholar
  36. Yoshikai, S., Sasaki, H., Dohura, K., Furuya, H., Sakaki, Y. (1990). Genomic organization of the human amyloid beta-protein precursor gene. Gene 87, 257–263.Google Scholar
  37. Yoshikai, S., Sasaki, H., Dohura, K., Furuya, H., Sakaki, Y. (1991). Genomic organization of the human amyloid beta-protein precursor gene. Gene 102, 291–292.Google Scholar
  38. Yoshioka, K., Miki, T., Katsuya, T., Ogihara, T., Sakaki, Y. (1991). The 717Val»Ile substitution in amyloid precursor protein is associated with familial Alzheimer's disease regardless of ethnic groups. Biochem. Biophys. Res. Commun. 178, 1141–1146.Google Scholar

Copyright information

© Springer-Verlag New York Inc 1993

Authors and Affiliations

  • Karen Rooke
    • 1
  • Chris Talbot
    • 3
  • Louise James
    • 2
  • Rakesh Anand
    • 2
  • John A. Hardy
    • 4
  • Alison M. Goate
    • 3
  1. 1.Department of Biochemistry and Molecular GeneticsSt. Mary's Hospital Medical SchoolLondonUK
  2. 2.Zeneca PharmaceuticalsMacclesfieldUK
  3. 3.Department of PsychiatryWashington University School of MedicineSt. LouisUSA
  4. 4.Suncoast Alzheimer's Disease Research Group, Department of PsychiatryUniversity of South FloridaTampaUSA

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