Presenilins are required for γ-secretase cleavage of β-APP and transmembrane cleavage of Notch-1 (original) (raw)

Nature Cell Biology volume 2, pages 463–465 (2000)Cite this article

Mutations in presenilin-1 (PS1) and presenilin-2 (PS2) cause early-onset familial Alzheimer’s disease1,2. The finding that presenilin mutations increase production of the amyloidogenic, 42-amino-acid form of amyloid β-protein (Aβ)3 gave rise to the proposal that presenilins function in the metabolism of the β-amyloid precursor protein (β-APP). That PS1 may have a direct function in the γ-secretase-mediated cleavage of β-APP that generates the Aβ carboxy terminus is indicated by the observations that Aβ production is reduced and accumulation of β-APP C-terminal fragments (CTFs) is increased in PS1-null cells4,5. Furthermore, dominant-negative aspartyl mutations in PS1 and PS2 also reduce Aβ production, and a combination of dominant-negative PS1 and PS2 mutations results in no detectable Aβ production at all6,7,8. A possible function of presenilins in Notch proteolytic processing and signalling has also been indicated by the facts that proteolytic release of the Notch-1 intracellular domain (NICD) is reduced in PS1-null cells9,10, and that developmental abnormalities consistent with reduced Notch signalling are observed in presenilin-deficient animal models7,9,10,11,12,13. Although these findings are consistent with the idea that presenilins are involved in γ-secretase cleavage and proteolytic release of NICD, the definitive test that they are absolutely required is whether production of Αβ and NICD is abolished in cells lacking both PS1 and PS2.

We analysed production of Aβ by immunoprecipitating it from the conditioned medium of blastocyst cultures, and then immunoblotting using a gel system that allows the 40- and 42-amino-acid forms to be distinguished17. PS1+/+PS2+/+ cells showed robust production of the 40-amino-acid form (Aβ40) and lower levels of Αβ42; the ratio of Aβ42:Aβ40 was 10–20% (Fig. 1b). Production of Αβ40 and Aβ42 was significantly reduced in PS1 –/–PS2+/+ and PS1 +/–_PS2_–/– cells, although significant residual Aβ production was observed ( Fig. 1b, d). In contrast, production of Aβ40 and Aβ42 was not detectable in media from two different _PS1_–/– _PS2_–/– cell lines (Fig. 1b, d). Titration of the medium showed that the assay could resolve <0.15% of the Αβ in PS1+/+PS2+/+ cells (<2 pM). Transfection with PS1 cDNA restored production of Aβ40 and Aβ42 in _PS1_–/–_PS2_–/– cells (Fig. 1c). The absence of Aβ generation in _PS1_–/–_PS2_–/– cells, together with the observed increase in accumulation of CTFs, indicates that presenilin expression is required for γ-secretase cleavage of β-APP.

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References

  1. Sherrington, R. et al. Nature 375, 754–760 (1995).
    Article CAS Google Scholar
  2. Levy-Lahad, E. et al. Science 269, 973–977 (1995).
    Article CAS Google Scholar
  3. Scheuner, D. et al. Nature Med. 2, 864– 870 (1996).
    Article CAS Google Scholar
  4. De Strooper, B. et al. Nature 398, 518–522 (1999).
    Article CAS Google Scholar
  5. Naruse, S. et al. Neuron 21, 1213–1221 (1998).
    Article CAS Google Scholar
  6. Wolfe, M.S. et al. Nature 398, 513–517 (1999).
    Article CAS Google Scholar
  7. Steiner, H. et al. J. Biol. Chem. 274, 28669– 28673 (1999).
    Article CAS Google Scholar
  8. Kimberly, W.T., Xia, W., Rahmati, T., Wolfe, M.S. and Selkoe, D.S. J. Biol. Chem. 275, 3173– 3178 (2000).
    Article CAS Google Scholar
  9. De Strooper, B. et al. Nature 398, 518–522 (1999).
    Article CAS Google Scholar
  10. Song, W. et al. Proc. Natl Acad. Sci. USA 96, 6959– 6963 (1999).
    Article CAS Google Scholar
  11. Levitan, D. & Greenwald, I. Nature 377, 351–355 (1995).
    Article CAS Google Scholar
  12. Shen, J. et al. Cell 89, 629–639 (1997).
    Article CAS Google Scholar
  13. Wong P. et al. Nature 387, 288–292 (1997).
  14. Struhl, G. & Greenwald, I. (1999). Nature 398, 522–525.
  15. Ye, Y., Lukhinova, N. & Fortini, M. E. Nature 398, 525– 529 (1999).
    Article CAS Google Scholar
  16. Donoviel, D. et al. Genes Dev. 13, 280 – 2810 (1999).
    Article Google Scholar
  17. Klafki, H. W., Wiltfang, J. & Staufenbiel, M. Analyt. Biochem. 237, 24– 29 (1996).
    Article CAS Google Scholar
  18. Schroeter, E. H., Kisslinger, J. A. & Kopan, R. Nature 393, 382– 386 (1998).
    Article CAS Google Scholar
  19. Herreman et al. Proc. Natl Acad. Sci. USA 96, 11872–11877 ( 1999).
  20. Yankner, B. A. Neuron 16, 921–932 ( 1996).
    Article CAS Google Scholar
  21. Busciglio, J., Gabuzda, D. H., Matsudaira, P. & Yankner, B. A. Proc. Natl Acad. Sci. USA 90,1993).

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Acknowledgements

This work was supported by grants from the NIH and a Zenith Award from the Alzheimer’s Association (to B.A.Y), by fellowships from the Medical Foundation (to Z.Z.) and from the Edward R. and Anne G. Lefler Foundation (to P.N), and by an NIH MRRC Core grant.

Correspondence and requests for materials should be addressed to B.A.Y.

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Author notes

  1. Zhuohua Zhang, Philip Nadeau, Weihong Song, Dorit Donoviel and Alan Bernstein: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Neurology, Harvard Medical School and Division of Neuroscience, Children’s Hospital, Boston, 02115, MA, USA
    Zhuohua Zhang, Philip Nadeau, Weihong Song, Menglan Yuan & Bruce A. Yankner
  2. Mount Sinai Hospital and Department of Medical Genetics and Microbiology, Samuel Lunenfeld Research Institute, University of Toronto, Toronto, M5S-1A8, Canada
    Dorit Donoviel & Alan Bernstein

Authors

  1. Zhuohua Zhang
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  2. Philip Nadeau
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  3. Weihong Song
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  4. Dorit Donoviel
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  5. Menglan Yuan
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  6. Alan Bernstein
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  7. Bruce A. Yankner
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Correspondence toBruce A. Yankner.

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Zhang, Z., Nadeau, P., Song, W. et al. Presenilins are required for γ-secretase cleavage of β-APP and transmembrane cleavage of Notch-1.Nat Cell Biol 2, 463–465 (2000). https://doi.org/10.1038/35017108

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