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The role of presenilin cofactors in the γ-secretase complex


Mutations in presenilin genes account for the majority of the cases of the familial form of Alzheimer's disease (FAD). Presenilin is essential for γ-secretase activity, a proteolytic activity involved in intramembrane cleavage of Notch and β-amyloid precursor protein (βAPP)1,2. Cleavage of βAPP by FAD mutant presenilin results in the overproduction of highly amyloidogenic amyloid β42 peptides3,4,5,6. γ-Secretase activity requires the formation of a stable, high-molecular-mass protein complex7,8,9,10,11 that, in addition to the endoproteolysed fragmented form of presenilin, contains essential cofactors including nicastrin12,13,14, APH-1 (refs 15–18) and PEN-2 (refs 16, 19). However, the role of each protein in complex formation and the generation of enzymatic activity is unclear. Here we show that Drosophila APH-1 (Aph-1) increases the stability of Drosophila presenilin (Psn) holoprotein in the complex. Depletion of PEN-2 by RNA interference prevents endoproteolysis of presenilin and promotes stabilization of the holoprotein in both Drosophila and mammalian cells, including primary neurons. Co-expression of Drosophila Pen-2 with Aph-1 and nicastrin increases the formation of Psn fragments as well as γ-secretase activity. Thus, APH-1 stabilizes the presenilin holoprotein in the complex, whereas PEN-2 is required for endoproteolytic processing of presenilin and conferring γ-secretase activity to the complex.

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Figure 1: Effects of Aph-1 on accumulation, stabilization and high-Mr complex formation of Psn holoprotein.
Figure 2: Role of Pen-2 in the metabolism of Psn and γ-secretase activity.
Figure 3: Role of PEN-2 in the metabolism of mammalian PS1.
Figure 4: Overexpression of Pen-2 in addition to Nct and Aph-1 increases the accumulation of Psn fragments and the γ-secretase activity.
Figure 5: Schematic depiction of the stepwise assembly and activation of the γ-secretase complex in Drosophila cells.

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We thank M. Miura, H. Kanuka, T. Igaki, K. Tei, H. Bito, M. Nakafuku and Y. Gotoh for suggestions on S2 and BG2 cells and RNAi assays; G. L. Boulianne, S. Cohen and Y. Ihara for providing Psn cDNA, anti-Smo antibody and anti-C4 antibody, respectively; Takeda Chemical Industries for continuous support to our studies; D.M.A. Mann for comments on the manuscript; and Y. Morohashi, N. Isoo, S. Tanaka and C. Sato for discussions and technical assistance. This work was supported by grants-in-aid from the Ministry of Health and Welfare and the Ministry of Education, Science, Culture and Sports for the 21st Century Center of Excellence programme, Japan. N.T. is a Research Fellow of the Japan Society for the Promotion of Science.

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Correspondence to Taisuke Tomita or Takeshi Iwatsubo.

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Takasugi, N., Tomita, T., Hayashi, I. et al. The role of presenilin cofactors in the γ-secretase complex. Nature 422, 438–441 (2003).

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