Inositol 1,4,5-Trisphosphate Receptor Is Located to the Inner Nuclear Membrane Vindicating Regulation of Nuclear Calcium Signaling by Inositol 1,4,5-Trisphosphate: DISCRETE DISTRIBUTION OF INOSITOL PHOSPHATE RECEPTORS TO INNER AND OUTER NUCLEAR MEMBRANES

Abstract

Transient rise in nuclear calcium concentration is implicated in the regulation of events controlling gene expression. Mechanism by which calcium is transported to the nucleus is vehemently debated. Inositol 1,4,5-trisphosphate (InsP3) and inositol-1,3,4,5-tetrakisphosphate (InsP4) receptors have been located to the nucleus and their role in nuclear calcium signaling has been proposed. Outer nuclear membrane was separated from the inner membrane. The two membrane preparations were, as best as possible, devoid of cross contamination as attested by marker enzyme activity, Western blotting with antilamin antibody, and electron microscopy. InsP4 receptor and Ca(2+)-ATPase were located to the outer nuclear membrane. InsP3 receptor was located to the inner nuclear membrane. ATP or InsP4 induced nuclear calcium uptake. External free calcium concentration, in the medium bathing the nuclei, determined the choice for ATP or InsP4-mediated calcium transport. We present a mechanistic model for nuclear calcium transport. According to this model, calcium can reach the nucleus envelope either by the action of ATP or InsP4. However, the calcium release from the nucleus envelope to the nucleoplasm is mediated by InsP3 through the activation of InsP3 receptor, which is located to the inner nuclear membrane. The action of InsP3 in this process was instantaneous and transient and was sensitive to heparin.