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The promoter region of the α-subunit of the calcium/calmodulin-dependent protein kinase II (α-CaMKII) gene was inserted into a β-galactosidase (β-gal) reporter plasmid, and β-gal activities were examined in neuroblastoma (NB2a) and pheochromocytoma (PC12) cells after transient or stable transfections. The α-CaMKII promoter was 12- to 45-fold more active in NB2a compared with PC12 cells after transient or stable transfections. All-trans retinoic acid (RA) stimulated reporter gene expression at both protein and mRNA levels in transfected PC12 cells. RA increased the level of endogenous α-CaMKII mRNA in untransfected PC12 cells by 4.4-fold. The transcription initiation site(s) (TIS) of the α-CaMKII gene in PC12 cells and rat brain was examined by RNase protection assays (RPA) and reverse transcriptase PCRs. The TIS for the α-CaMKII/β-gal reporter gene in transfected PC12 cells was indistinguishable from the TIS+1 in rat hippocampus. In contrast, the only detectable TIS for the α-CaMKII gene in untransfected PC12 cells was located near the ATG translation start codon, 147 nucleotides 3′ to TIS+1 in hippocampus. This unusual TIS was also the predominant TIS in rat cerebellum. These results suggest that the α-CaMKII promoter may contain sequences that respond directly or indirectly to RA. In addition, the unusual TIS of the α-CaMKII gene in PC12 cells and rat cerebellum may contribute to the very low expression of this gene compared with that in hippocampus.

Retinoic Acid Stimulates α-CAMKII Gene Expression in PC12 Cells at a Distinct Transcription Initiation Site