Zinc regulation of transcriptional activity during retinoic acid-induced neuronal differentiation

Zinc deficiency impairs the proliferation and differentiation of stem cells in the central nervous system that participate in neurogenesis. To examine the molecular mechanisms responsible for the role of this essential nutrient in neuronal precursor cells and neuronal differentiation, we identified zinc-dependent changes in the DNA-binding activity of zinc finger proteins and other transcription factors in proliferating human Ntera-2 neuronal precursor cells undergoing retinoic acid-stimulated differentiation into a neuronal phenotype. We found that zinc deficiency altered binding activity of 28 transcription factors including retinoid X receptor (RXR) known to participate in neuronal differentiation. Alterations in zinc finger transcription factor activity were not simply the result of removal of zinc from these proteins during zinc deficiency, as the activity of other zinc-binding transcription factors such as the glucocorticoid receptor was increased by as much as twofold over zinc-adequate conditions, and nonzinc-binding transcription factors such as nuclear factor-1 and heat shock transcription factor-1 were increased by as much as fourfold over control. Western analysis did not detect significant decreases in total RXR protein abundance in neuronal precursors, suggesting that the decrease in DNA-binding activity was not simply the result of a reduction in RXR levels in neuronal precursor cells. Rather, use of a reporter gene construct containing retinoic acid response elements upstream from a luciferase coding sequence revealed that zinc deficiency results in decreased transcriptional activity of RXR and reductions in retinoic acid-mediated gene transcription during neuronal differentiation. These results show that zinc deficiency has implications for both developmental and adult neurogenesis.