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Upregulation of BiP and CHOP by the unfolded-protein response is independent of presenilin expression

Nature Cell Biology volume 2, pages 863–870 (2000)Cite this article

Abstract

Presenilin 1 (PS1), a polytopic membrane protein, has a critical role in the trafficking and proteolysis of a selected set of transmembrane proteins. The vast majority of individuals affected with early onset familial Alzheimer's disease (FAD) carry missense mutations in PS1. Two studies have suggested that loss of PS1 function, or expression of FAD-linked PS1 variants, compromises the mammalian unfolded-protein response (UPR), and we sought to evaluate the potential role of PS1 in the mammalian UPR. Here we show that that neither the endoplasmic reticulum (ER) stress-induced accumulation of BiP and CHOP messenger RNA, nor the activation of ER stress kinases IRE1α and PERK, is compromised in cells lacking both PS1 and PS2 or in cells expressing FAD-linked PS1 variants. We also show that the levels of BiP are not significantly different in the brains of individuals with sporadic Alzheimer's disease or PS1-mediated FAD to levels in control brains. Our findings provide evidence that neither loss of PS1 and PS2 function, nor expression of PS1 variants, has a discernable impact on ER stress-mediated induction of the several established `readouts' of the UPR pathway.

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Figure 1: UPR-mediated induction of BiP and CHOP expression in primary PS1−/− and PS1+/− fibroblasts treated with tunicamycin.
Figure 2: ER stress signalling and the induction of BiP and CHOP mRNAs in PS1−/−/PS2−/− double-null cells.
Figure 3: Expression of the FAD-linked PS1 C410Y variant does not affect the UPR in stable N2a cell lines.
Figure 4: Kinetic analysis of BiP and CHOP induction in tunicamycin-treated stable N2a lines.
Figure 5: Analysis of ER stress-mediated BiP induction in stable pools of human 293 cells.
Figure 6: Western blot analysis of BiP and GRP94 induction in stable 293 cells.
Figure 7: Western blot analysis of BiP protein levels in brains of FAD patients and mutant PS1 transgenic mice.

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Acknowledgements

This work was supported by NIH 1PO1 grant AG14248 (S.S.S.), NIEHS grant ES08681 (D.R), NIH, NCI grant CA27607 (A.S.L), the Alzheimer's Association (G.T), the Adler Foundation (G.T. and J.Y.L), and by an award to the University of Chicago's Division of Biological Sciences under the Research Resources Program for Medical Schools of the Howard Hughes Medical Institute (G.T.). N.S. and F.U are supported by a research fellowship from the Japan Society for the Promotion of Science and grants-in-aid from the Ministry of Education, Science, Culture and Sports, Japan. We thank C. A. Saura, T. Tomita and T. Iwatsubo for Aβ analysis; D. R. Borchelt for transgenic mouse tissue; J. Troncoso for control and sporadic AD brain tissue; and L. Hendriks and C. V. Broeckhoven for FAD brain tissue.

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Authors and Affiliations

  1. Department of Neurobiology, Pharmacology and Physiology The University of Chicago, Knapp R212, 924 East 57th street , Chicago, 60637, Illinois, USA

    Naoyuki Sato, Jae Yoon Leem, Seong-Hun Kim, Margaret L. Veselits, Sangram S. Sisodia & Gopal Thinakaran

  2. Skirball Institute of Biomolecular Medicine, New York University Medical School, New York, 10016, New York, USA

    Fumihiko Urano & David Ron

  3. Department of Biochemistry and Molecular Biology, USC/Norris Comprehensive Cancer Center, University of Southern California Keck School of Medicine, Los Angeles, 90089 , California, USA

    Mingqing Li & Amy S. Lee

  4. Department of Molecular and Medical Genetics, Samuel Lunenfeld Research Institute of Mount Sinai Hospital, University of Toronto, Toronto, M5S-1A8, Ontario, Canada

    Dorit Donoviel & Alan Bernstein

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Correspondence to Gopal Thinakaran.

Supplementary information

Figure S1

Semi-quantitative RT-PCR analysis of BiP and CHOP mRNA levels in PS1-/- and PS1+/- fibroblasts. (PDF 224 kb)

Figure S2 Activation of the UPR is unimpaired by the expression of a loss of function PS1 mutant.

Figure S3 Kinetic analysis of BiP and CHOP mRNA in N2a cell lines expressing Wt PS1 or PS1 D385A mutant.

Figure S4 Coomassie blue staining of brain homogenates of controls, FAD patients and sporadic AD patients.

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Sato, N., Urano, F., Yoon Leem, J. et al. Upregulation of BiP and CHOP by the unfolded-protein response is independent of presenilin expression. Nat Cell Biol 2, 863–870 (2000). https://doi.org/10.1038/35046500

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