Dr. F. Cambi is Department of Neurology, Thomas Jefferson Medical School, Philadelphia, PA 19017, U.S.A.
5′ Flanking DNA Sequences Direct Cell-Specific Expression of Rat Tyrosine Hydroxylase
Version of Record online: 5 OCT 2006
Journal of Neurochemistry
Volume 53, Issue 5, pages 1656–1659, November 1989
How to Cite
Cambi, F., Fung, B. and Chikaraishi, D. (1989), 5′ Flanking DNA Sequences Direct Cell-Specific Expression of Rat Tyrosine Hydroxylase. Journal of Neurochemistry, 53: 1656–1659. doi: 10.1111/j.1471-4159.1989.tb08567.x
- Issue online: 5 OCT 2006
- Version of Record online: 5 OCT 2006
- Received August 9, 1989; accepted August 14, 1989.
- Tyrosine hydroxylase;
- Binding factor API;
- Cyclic AMP response element
Abstract: Tyrosine hydroxylase (TH) is selectively expressed in cat-echolaminergic neurons and in chromaffin cells of the adrenal medulla. Constructs in which 5’flanking sequences of the rat TH gene directed expression of bacterial chloramphenicol acetyltransferase (CAT) were transfected into cell lines and assayed for transient expression of CAT. In most nonexpressing cell lines, CAT levels were < 5% of that found in a TH-positive pheochromocytoma line (PC8b). In two lines described here, a rat anterior pituitary (pell line (GH4) and a rat fibroblast line (Fr3T3), CAT expression reached 12 and 20%, respectively, of the PC8b level. Greater than 90% of the PC8b activity was lost when sequences between -212 and - 187 (in relation to the transcriptional initiation site) were deleted. Further deletions that removed the cyclic AMP response element (CRE)|(-45) and the TATA box at -29 reduced transcriptional activity to background in all three lines. These data suggest that 212 nucleotides of the 5’sequence are sufficient for pheochromocytoma expression and that information between —212 and —187, which includes an API site (-206 to —200), is essential for full transcriptional activity. In addition, sites for other protein transcription factors (AP2, POU/Oct, SP1, and CRE) reside between -221 and -38 and are largely conserved between the human and rat gene.