Novel transcriptional mechanisms are involved in regulating preproenkephalin gene expression in vivo.

For the dissection of the temporal and spatial patterns of cell- and tissue-specific gene expression an understanding of the contributing regulating mechanisms is required. We now confirm that there are novel mechanisms regulating preproenkephalin gene expression in basal as well as cholinergic agonist treated rats. Moreover, we demonstrate that these novel transcriptional mechanisms are consistent with RNA intragenic elongation pausing, alternate promoter usage, and small sense and antisense RNA transcription from the preproenkephalin gene locus. We report that while basal striatal and olfactory bulb proenkephalin RNA transcripts are initiated from the "normal" proximal promoter, in cerebellum de novo RNA transcription appears to be initiated from the distal so-called "germ-cell" promoter. Furthermore, "normally" initiated olfactory bulb proenkephalin RNA transcripts appear to be down-regulated by the time the RNA polymerase II complex reaches the first preproenkephalin intron, in a way that is consistent with RNA elongation pausing. As the pattern of small sense and antisense transcripts found associated with this gene's expression is tissue-specific, we suggest that they may also play a role in regulating gene expression. The understanding of this gene's regulation should have widespread importance, not only to those interested in opioid gene expression, but also to those interested in gene regulation, in general.