Depletion of Intracellular Ca2+ by Caffeine and Ryanodine Induces Apoptosis of Chinese Hamster Ovary Cells Transfected with Ryanodine Receptor

Pan, Zui; Damron, Derek S.; Nieminen, Anna‐Liisa; Bhat, Manjunatha B.; Ma, Jianjie

doi:10.1074/jbc.m908329199

Cited by 89 publications

(73 citation statements)

References 44 publications

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Section: Methodsmentioning

confidence: 99%

“…The GFP-RyR construct was obtained by replacing the first 290 amino acids of the rabbit skeletal muscle RyR with GFP (16). The E4032A RyR mutant was generated using site-directed mutagenesis, as described in previous studies (14).Cell Culture, Transfection, and DNA Laddering-c1148 and mg-5 are stable clones of Chinese hamster ovary (CHO) cells permanently expressing the RyR and MG29 proteins, respectively. The cells were grown at 37°C and 5% CO 2 in Ham's F-12 medium supplemented with 10% fetal bovine serum, 100 units/ml penicillin, 100 g/ml streptomycin, and 0.5 mg/ml G418.…”

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confidence: 99%

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Co-expression of MG29 and Ryanodine Receptor Leads to Apoptotic Cell Death

Pan¹,

Hirata²,

Nagaraj³

et al. 2004

Journal of Biological Chemistry

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Perturbation of intracellular Ca2؉ homeostasis has been shown to regulate the process of cell proliferation and apoptosis. Our previous studies show that mitsugumin 29 (MG29), a synaptophysin-related protein localized in the triad junction of skeletal muscle, serves an essential role in muscle Ca 2؉ signaling by regulating the process of store-operated Ca 2؉ entry. Here we report a functional interaction between MG29 and the ryanodine receptor (RyR)/Ca 2؉ release channel. The purified MG29 protein enhances activity of the RyR/Ca 2؉ release channel incorporated into the lipid bilayer membrane. Coexpression of MG29 and RyR in Chinese hamster ovary cells leads to apoptotic cell death resulting from depletion of intracellular Ca 2؉ stores, despite neither protein expression alone exhibits any significant effect on cell viability. In transient expression studies, the presence of RyR in the endoplasmic reticulum leads to retention of MG29 from the plasma membrane into the intracellular organelles. This functional interaction between MG29 and RyR could have important implications in the Ca 2؉ signaling processes of muscle cells. Our data also show that perturbation of intracellular Ca 2؉ homeostasis can serve as a key signal in the initiation of apoptosis. Ca2ϩ as a second messenger plays important roles in a variety of cellular processes, ranging from cell motility to enzymatic control, and to gene regulation and apoptosis. The ryanodine receptors (RyR) 1 /Ca 2ϩ release channels are present in the endo/(sarco)plasmic reticulum of virtually every cell types, providing a pathway for fast Ca 2ϩ release from intracellular stores (1-4). Mitsugumin29 (MG29) is a novel membrane protein localized specifically to the triad junction of skeletal muscle (5). The primary amino acid sequence of MG29 shares ϳ45% homology to synaptophysin, a family of proteins with presumed roles in secretion and neurotransmitter release (6, 7). The MG29 protein appears to participate in the excitation-contraction coupling process of skeletal muscle, since mutant mice lacking the MG29 gene exhibit apparent reduction in contractile force, with altered structure of the triad junction (8), and increased susceptibility to fatigue of the skeletal muscle (9, 10).Several studies have suggested a central role for the participation of endoplasmic reticulum (ER) and intracellular Ca 2ϩ release in the initiation of apoptosis (11-13). Our previous studies have shown that depletion of intracellular Ca 2ϩ stores via activation of the RyR/Ca 2ϩ release channel could induce apoptosis (14, 15). Here we tested the hypothesis that overexpression of MG29 can influence the intracellular Ca 2ϩ homeostasis through changes of the RyR/Ca 2ϩ release function. We show that co-expression of MG29 and RyR in cultured cells leads to apoptotic cell death resulting from depletion of intracellular Ca 2ϩ stores, and the purified MG29 protein can enhance activity of the RyR/Ca 2ϩ release channel incorporated into the lipid bilayer membrane. Our data provide evidence for a functional ...

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Co-expression of MG29 and Ryanodine Receptor Leads to Apoptotic Cell Death

Pan¹,

Hirata²,

Nagaraj³

et al. 2004

Journal of Biological Chemistry

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“…1,10,11 In contrast to Bcl-2, 12 ± 14 no local ER effects have been described for Bcl-X L . 15 Viral mitochondrial inhibitor of apoptosis (vMIA) is encoded by the Cytomegalovirus UL37 gene, has no obvious structural similarity with Bcl-2-like molecules, and is exclusively found in mitochondrial membranes, where it specifically interacts with ANT but not with VDAC, as shown by mass spectroscopic identification of vMIAinteracting proteins and confirmed by co-immunoprecipitation assays. 16 ± 18 TM, BFA, and TG induced signs of MMP in several different cell lines including B cell lymphoma BJAB ( Figure 1a) and cervical carcinoma HeLa cells (Figure 1b).…”

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confidence: 99%

Endoplasmic reticulum stress-induced cell death requires mitochondrial membrane permeabilization

et al. 2002

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“…6 Studies have shown that Bax and other proapoptotic members of the Bcl-2 family could modulate Ca 2 þ stores in the endoplasmic reticulum (ER) and change mitochondrial matrix Ca 2 þ (Ca m 2 þ ) contents. 7,8 In addition, mitochondrial membrane integrity is altered by excessive Ca 2 þ uptake. 9 Bax and another multidomain protein, Bak, may translocate or localize in the ER, deplete Ca 2 þ from the ER and activate caspase-12.…”

Section: Introductionmentioning

confidence: 99%

“…Some of these alterations include changes in Ca m 2 þ uptake promoted by apoptotic stimuli, which increase the probability of PTP activation. 16 Depletion of intracellular Ca 2 þ stores has also been related to the induction of apoptosis, 17 and a decrease in Ca 2 þ concentration within the ER, leading to Ca m 2 þ accumulation, determines the mode and amount of cytochrome c release 18 and apoptosis. 19 On the other hand, recent studies have demonstrated that Bax and Bak deficiency causes a decrease in Ca 2 þ at the ER level and a decrease in mitochondrial Ca 2 þ uptake.…”

Section: Introductionmentioning

confidence: 99%

Bax affects intracellular Ca2+ stores and induces Ca2+ wave propagation

et al. 2004

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In the present study, we evaluated proapoptotic protein Bax on mitochondria and Ca 2 þ homeostasis in primary cultured astrocytes. We found that recombinant Bax (rBax, 10 and 100 ng/ml) induces a loss in mitochondrial membrane potential (DW m ). This effect might be related to the inhibition of respiratory rates and a partial release of cytochrome c, which may change mitochondrial morphology. The loss of DW m and a selective permeabilization of mitochondrial membranes contribute to the release of Ca 2 þ from the mitochondria. This was inhibited by cyclosporin A (5 lM) and Ruthenium Red (1 lg/ml), indicating the involvement of mitochondrial Ca 2 þ transport mechanisms. Bax-induced mitochondrial Ca 2 þ release evokes Ca 2 þ waves and wave propagation between cells. Our results show that Bax induces mitochondrial alteration that affects Ca 2 þ homeostasis and signaling. These changes show that Ca 2 þ signals might be correlated with the proapoptotic activities of Bax.

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Depletion of Intracellular Ca2+ by Caffeine and Ryanodine Induces Apoptosis of Chinese Hamster Ovary Cells Transfected with Ryanodine Receptor

Cited by 89 publications

References 44 publications

Co-expression of MG29 and Ryanodine Receptor Leads to Apoptotic Cell Death

Co-expression of MG29 and Ryanodine Receptor Leads to Apoptotic Cell Death

Endoplasmic reticulum stress-induced cell death requires mitochondrial membrane permeabilization

Bax affects intracellular Ca2+ stores and induces Ca2+ wave propagation

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