Contribution of presenilin/γ-secretase to calsenilin-mediated apoptosis

doi:10.1016/S0006-291X(03)00688-0

Biochemical and Biophysical Research Communications

Volume 305, Issue 1, 23 May 2003, Pages 62-66

https://doi.org/10.1016/S0006-291X(03)00688-0 Get rights and content

Abstract

Mutant presenilins cause early-onset of familial Alzheimer’s disease and render cells vulnerable to apoptosis. Calsenilin/DREAM/KChIP3 is a multifunctional calcium-binding protein that interacts with presenilin and mediates calcium-mediated apoptosis. In the present study, we report that the calsenilin-mediated apoptosis is regulated by presenilin. The expression of calsenilin was highly up-regulated in neuronal cells undergoing Aβ42-triggered cell death. The incidence of calsenilin-mediated apoptosis was diminished in presenilin-1^−/− mouse embryonic fibroblast cells or neuronal cells stably expressing a loss-of-function presenilin-1 mutant. On the contrary, an array of familial Alzheimer’s disease-associated presenilin mutants (gain-of-function) increased calsenilin-induced cell death. Moreover, γ-secretase inhibitors, including compound E and DAPT, decreased the calsenilin-induced cell death. These results suggest that the pro-apoptotic activity of calsenilin coordinates with presenilin/γ-secretase activity to play a crucial role in the neuronal death of Alzheimer’s disease.

Section snippets

Materials and methods

Cell cultures and preparation of Aβpeptide. B103 rat neuroblastoma cells, SK-N-BE2(c) human neuroblastoma cells, and mouse embryonic fibroblasts (MEFs) from wild-type or PS1-null mice [19] were grown in DMEM (Gibco-BRL) supplemented with 10% heat-inactivated fetal bovine serum. PS1^−/− or PS1^+/+ MEFs at the passage between four and seven were used in the experiments. SK-N-BE2(c) cells stably expressing wild-type PS1 or PS2, PS1-M146V, PS2-N141I, or empty plasmid (pcDNA3); B103 cells expressing

Results and discussion

The accumulation of Aβ in the brains of AD patients has been implicated as a cause of the neuronal loss that occurs in AD. Previously, we have shown that the pro-apoptotic activity of calsenilin mediated calcium-signal-triggered cell death [18]. We also observed that calsenilin was highly up-regulated in the brains of AD patients and AD transgenic mice (manuscript in preparation). However, the apoptotic signaling pathway mediated by calsenilin has not been fully elucidated. To determine the

Acknowledgements

We thank Dr. M. Paul Murphy for the gift of compound E and are grateful to Dr. De Strooper for providing PS1^−/− MEF cell. This work was supported by the grant from National Research Laboratory program (to Y. Jung) and Life Phenomena and Function Research Group Program (2000) of the Korean MOST.

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Cited by (27)

CLN3, at the crossroads of endocytic trafficking
2021, Neuroscience Letters
The CLN3 gene was identified over two decades ago, but the primary function of the CLN3 protein remains unknown. Recessive inheritance of loss of function mutations in CLN3 are responsible for juvenile neuronal ceroid lipofuscinosis (Batten disease, or CLN3 disease), a fatal childhood onset neurodegenerative disease causing vision loss, seizures, progressive dementia, motor function loss and premature death. CLN3 is a multipass transmembrane protein that primarily localizes to endosomes and lysosomes. Defects in endocytosis, autophagy, and lysosomal function are common findings in CLN3-deficiency model systems. However, the molecular mechanisms underlying these defects have not yet been fully elucidated. In this mini-review, we will summarize the current understanding of the CLN3 protein interaction network and discuss how this knowledge is starting to delineate the molecular pathogenesis of CLN3 disease. Accumulating evidence strongly points towards CLN3 playing a role in regulation of the cytoskeleton and cytoskeletal associated proteins to tether cellular membranes, regulation of membrane complexes such as channels/transporters, and modulating the function of small GTPases to effectively mediate vesicular movement and membrane dynamics.
Modulation of Kv4.2/KChIP3 interaction by the ceroid lipofuscinosis neuronal 3 protein CLN3
2020, Journal of Biological Chemistry
Citation Excerpt :
Because of its interaction with presenilin, KChIP3 functions as a Ca2+ sensor for the γ-secretase, which mediates enhanced enzyme activity at elevated Ca2+ levels (31). The presenilin/γ-secretase enzyme complex is integral to the proapoptotic activity of KChIP3 (32). The intriguing finding that KChIP3 can also interact with CLN3 (23) may add another aspect to the involvement of KChIP3 in brain pathophysiology, because mutated CLN3 is found in JNCL (24).
Voltage-gated potassium (Kv) channels of the Kv4 subfamily associate with Kv channel–interacting proteins (KChIPs), which leads to enhanced surface expression and shapes the inactivation gating of these channels. KChIP3 has been reported to also interact with the late endosomal/lysosomal membrane glycoprotein CLN3 (ceroid lipofuscinosis neuronal 3), which is modified because of gene mutation in juvenile neuronal ceroid lipofuscinosis (JNCL). The present study was undertaken to find out whether and how CLN3, by its interaction with KChIP3, may indirectly modulate Kv4.2 channel expression and function. To this end, we expressed KChIP3 and CLN3, either individually or simultaneously, together with Kv4.2 in HEK 293 cells. We performed co-immunoprecipitation experiments and found a lower amount of KChIP3 bound to Kv4.2 in the presence of CLN3. In whole-cell patch-clamp experiments, we examined the effects of CLN3 co-expression on the KChIP3-mediated modulation of Kv4.2 channels. Simultaneous co-expression of CLN3 and KChIP3 with Kv4.2 resulted in a suppression of the typical KChIP3-mediated modulation; i.e. we observed less increase in current density, less slowing of macroscopic current decay, less acceleration of recovery from inactivation, and a less positively shifted voltage dependence of steady-state inactivation. The suppression of the KChIP3-mediated modulation of Kv4.2 channels was weaker for the JNCL-related missense mutant CLN3_R334C and for a JNCL-related C-terminal deletion mutant (CLN3ΔC). Our data support the notion that CLN3 is involved in Kv4.2/KChIP3 somatodendritic A-type channel formation, trafficking, and function, a feature that may be lost in JNCL.
The regulation of apoptosis by the downstream regulatory element antagonist modulator/potassium channel interacting protein 3 (DREAM/KChIP3) through interactions with hexokinase I
2013, Biochemical and Biophysical Research Communications
Citation Excerpt :
Dream−/− mice display reduced responses to pain, and improved contextual fear conditioning [13,14]. DREAM regulates apoptosis [15,16] by altering intracellular Ca2+ signaling [16], changing presenilin 1 and 2 activity [9,15,17] and by binding to peroxiredoxin 3, an antioxidant enzyme [18]. DREAM binds to the carboxyl-terminal portion of presenilin to regulate γ-secretase function [9].
The EF-hand protein, DREAM/KChIP3 (henceforth referred to as DREAM), regulates apoptosis by incompletely understood mechanisms. We demonstrate that in the presence of Ca²⁺, DREAM interacts with hexokinase I, a protein known to bind mitochondria and regulate apoptosis. A mutant DREAM protein construct incapable of binding Ca²⁺ does not associate with hexokinase I. The amino-terminal portion of DREAM is required for binding to hexokinase I, as a DREAM construct lacking the first 94 amino terminal residues fails to bind hexokinase I. Expression of DREAM in neuroblastoma cells enhances cisplatin mediated caspase-3 activity. Simultaneous expression of hexokinase I in such cells reduces DREAM-stimulated apoptosis. DREAM overexpression in neuroblastoma cells reduces hexokinase I localization on isolated mitochondria. The interaction of DREAM with hexokinase I may be important in the regulation of neuronal apoptosis.
Secretases as therapeutic targets for Alzheimer's disease
2011, Biochemical and Biophysical Research Communications
Citation Excerpt :
Assembly of the four components of γ-secretase leads to PS autoproteolysis into two fragments that contribute an aspartate to the active site. Much evidence suggests that PS or γ-secretase is also involved in the regulation of cellular Ca2+ homeostasis and Ca2+-mediated cell death [33,34]. NCT undergoes complex N-glycosylation during its maturation within the ER and Golgi compartments [35].
Accumulation of amyloid-β (Aβ) is widely accepted as the key instigator of Alzheimer’s disease (AD). The proposed mechanism is that accumulation of Aβ results in inflammatory responses, oxidative damages, neurofibrillary tangles and, subsequently, neuronal/synaptic dysfunction and neuronal loss. Given the critical role of Aβ in the disease process, the proteases that produce this peptide are obvious targets. The goal would be to develop drugs that can inhibit the activity of these targets. Protease inhibitors have proved very effective for treating other disorders such as AIDS and hypertension. Mutations in APP (amyloid-β precursor protein), which flanks the Aβ sequence, cause early-onset familial AD, and evidence has pointed to the APP-to-Aβ conversion as a possible therapeutic target. Therapies aimed at modifying Aβ-related processes aim higher up the cascade and are therefore more likely to be able to alter the progression of the disease. However, it is not yet fully known whether the increases in Aβ levels are merely a result of earlier events that were already causing the disease.
Contribution of γ-secretase to calcium-mediated cell death
2010, Neuroscience Letters
Presenilins are the catalytic subunit of the large γ-secretase complex, that promotes intramembranous proteolysis of the beta-amyloid precursor protein (APP), resulting in the production of beta-amyloid (Aβ). Mutant presenilin causes early-onset familial Alzheimer's disease (FAD), is related to abnormal Ca²⁺ signaling, and render cells vulnerable to cell death. In the present study, we demonstrated that Ca²⁺-mediated cell death is functionally associated with γ-secretase activity. We found that γ-secretase activity was elevated during Ca²⁺-mediated cell death. Using selective γ-secretase inhibitors, we examined the role of γ-secretase in cell death triggered by increased intracellular Ca²⁺. Indeed, treatment with the selective γ-secretase inhibitors, compound E, DAPT, or L-685.458 significantly decreased Ca²⁺-triggered cell death with that of the controls, but did not affect staurosporin or tunicamycin-mediated cell death. These results implicate the role of γ-secretase activity in Ca²⁺-mediated cell death.
An RNA aptamer that recognizes a specific conformation of the protein calsenilin
2007, Bioorganic and Medicinal Chemistry
The generation of molecules that selectively recognize specific conformations of a protein is an important component of the elucidation protein function. We have used SELEX (Systematic Evolution of Ligands by EXponential enrichment) technology to produce aptamers that bind in a conformationally selective manner to calsenilin, which involved in Ca²⁺-mediated apoptotic signaling. Since the conformations of calsenilin are quite different in the presence and absence of Ca²⁺, aptamers were selected against the dimeric protein both under calcium-bound and calcium-free conditions. We have found that aptamer-12 selectively binds to the dimeric form of the protein in the presence of calcium ion, while the binding of aptamer-2 does not discriminate between the Ca²⁺ bound and unbound protein. Data obtained from biochemical and biophysical experiments suggest that a dominant conformation of calcium-bound calsenilin exists in one dominant conformation and that one aptamer can be generated to recognize this conformation. In addition, observation made in this effort that aptamers selected against the two different conformations of calsenilin have different characteristics suggest that aptamers can serve as a plausible tool for recognizing various conformations of proteins, even those caused by interactions with small molecules or ions such as Ca²⁺.

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Biochemical and Biophysical Research Communications

Abstract

Section snippets

Materials and methods

Results and discussion

Acknowledgements

References (25)

Involvement of caspases in proteolytic cleavage of Alzheimer’s amyloid-rbeta precursor protein and amyloidogenic A β peptide formation

Cell

FEBS Lett.

Requirement of the familial Alzheimer’s disease gene PS2 for apoptosis. Opposing effect of ALG-3

J. Biol. Chem.

Presenilin-1 and -2 are molecular targets for γ-secretase inhibitors

J. Biol. Chem.

Mutant genes in familial Alzheimer’s disease and transgenic models

Anuu. Rev. Neurosci.

Apoptosis is induced by β-amyloid in cultured central nervous system neurons

Proc. Natl. Acad. Sci. USA

Aggregated amyloid-β protein induces cortical neuronal apoptosis and concomitant apoptotic pattern of gene induction

J. Neurosci.

Increased production of 4 kDa amyloid β peptide in serum deprived human primary neuron cultures: possible involvement of apoptosis

J. Neurosci.

Apoptosis in the nervous system

Nature

Alzheimer’s PS-1 mutation perturbs calcium homeostasis and sensitizes PC12 cells to death induced by amyloid β-peptide

NeuroReport

Interfering with apoptosis: Ca²⁺-binding protein ALG-2 and Alzheimer’s disease gene ALG-3

Science

Participation of presenilin 2 in apoptosis: enhanced basal activity conferred by an Alzheimer mutation

Science

Cited by (27)

CLN3, at the crossroads of endocytic trafficking

Modulation of Kv4.2/KChIP3 interaction by the ceroid lipofuscinosis neuronal 3 protein CLN3

The regulation of apoptosis by the downstream regulatory element antagonist modulator/potassium channel interacting protein 3 (DREAM/KChIP3) through interactions with hexokinase I

Secretases as therapeutic targets for Alzheimer's disease

Contribution of γ-secretase to calcium-mediated cell death

An RNA aptamer that recognizes a specific conformation of the protein calsenilin

Contribution of presenilin/γ-secretase to calsenilin-mediated apoptosis

Abstract

Section snippets

Materials and methods

Results and discussion

Acknowledgements

Cell

FEBS Lett.

J. Biol. Chem.

J. Biol. Chem.

Mutant genes in familial Alzheimer’s disease and transgenic models

Anuu. Rev. Neurosci.

Apoptosis is induced by β-amyloid in cultured central nervous system neurons

Proc. Natl. Acad. Sci. USA

Aggregated amyloid-β protein induces cortical neuronal apoptosis and concomitant apoptotic pattern of gene induction

J. Neurosci.

Increased production of 4 kDa amyloid β peptide in serum deprived human primary neuron cultures: possible involvement of apoptosis

J. Neurosci.

Apoptosis in the nervous system

Nature

Alzheimer’s PS-1 mutation perturbs calcium homeostasis and sensitizes PC12 cells to death induced by amyloid β-peptide

NeuroReport

Interfering with apoptosis: Ca2+-binding protein ALG-2 and Alzheimer’s disease gene ALG-3

Science

Participation of presenilin 2 in apoptosis: enhanced basal activity conferred by an Alzheimer mutation

Science

Interfering with apoptosis: Ca²⁺-binding protein ALG-2 and Alzheimer’s disease gene ALG-3