Presenilin-2 dampens intracellular Ca²⁺ stores by increasing Ca²⁺ leakage and reducing Ca²⁺ uptake

We have previously shown that familial Alzheimer’s disease mutants of presenilin-2 (PS2) and, to a lesser extent, of presenilin-1 (PS1) lower the Ca²⁺ concentration of intracellular stores. We here examined the mechanism by which wild-type and mutant PS2 affect store Ca²⁺ handling. By using HeLa, SH-SY5Y and MEFs as model cells, and recombinant aequorins as Ca²⁺ probes, we show evidence that transient expression of either wild-type or mutant PS2 increases the passive Ca²⁺ leakage: both ryanodine- and IP₃-receptors contribute to Ca²⁺ exit out of the ER, whereas the ribosome translocon complex is not involved. In SH-SY5Y cells and MEFs, wild-type and mutant PS2 potently reduce the uptake of Ca²⁺ inside the stores, an effect that can be counteracted by over-expression of SERCA-2B. On this line, in wild-type MEFs, lowering the endogenous level of PS2 by RNA interference, increases the Ca²⁺-loading capability of intracellular stores. Furthermore, we show that in PS double knockout MEFs, reduction of Ca²⁺ stores is mimicked by the expression of PS2-D366A, a loss-of-function mutant, uncleaved because also devoid of presenilinase activity but not by co-expression of the two catalytic active fragments of PS2. In summary, both physiological and increased levels of wild-type and mutant PS2 reduce the Ca²⁺ uptake by intracellular stores. To exert this newly described function, PS2 needs to be in its full-length form, even if it can subsequently be cleaved.