Abstract
Magnesium transporter subtype 1 (MagT1) is a newly discovered and evolutionarily conservative magnesium membrane transporter with channel like properties. Previous reports have demonstrated that MagT1 is important to cellular magnesium homeostasis. In this study, we investigated whether drosophila MagT1 (dMagT1) was functionally regulated by PKC activation in vitro. With patch clamping, we have observed that whole cell currents of wild type dMagT1 were magnesium selective and non-voltage dependent when expressed in a human neuroblastoma SH-SY5Y cell line. Furthermore, dMagT1 currents were significantly increased in cells treated with a non specific PKC activator PMA, but not in cells treated with the inactive form of PMA, 4α-PMA. Lastly, we have demonstrated that upregulation of dMagT1 currents by PKC activation involves specific PKC phorsphorylation sites in dMagT1. Of all three dMagT1 mutants created for testing the putative PKC phorsphorylation sites, dMagT1-S35A displayed a significant increase of whole cell currents while dMagT1-S100A and -S108A were not affected by PKC activation. Thus, we have demonstrated that dMagT1 is a magnesium selective transporter with basic biophysical characters similar to its mammalian homolog and can be functionally upregulated by PKC activation. Both dMagT1 Ser100 and Ser106 are equally important to this PKC-dependent modulation, therefore the most likely molecular sites for PKC phorsphorylation. The data presented here may establish a general regulatory mechanism for MagT1 by PKC activation.
