Analysis of the neuronal promoter of the rat aromatic L-amino acid decarboxylase gene.
The rat aromatic L-amino acid decarboxylase (AADC) gene contains alternative promoters directing expression of neuronal and nonneuronal mRNAs that differ only in their 5' untranslated regions (UTRs). We have analyzed the expression of the neuronal promoter of the AADC gene in cells synthesizing catecholamines and serotonin, as well as in non-AADC-expressing cells. We demonstrate the use of the neuronal-specific UTR in individual dopamine-, norepinephrine-, and serotonin-containing neurons. Transfection analyses show that the rat AADC neuronal promoter, containing 2,400 bp upstream of the transcription start site and including the 68-bp untranslated exon 2, can activate transcription from a reporter gene in both catecholaminergic and serotonergic cell lines. These analyses identified several positive and negative cis-active elements within this region. Unexpectedly, we observed that this promoter, when removed from its native context within the AADC gene, can also direct expression of a reporter gene in cells that do not normally express AADC mRNA. These results suggest that tissue-specific expression of the neuronal promoter may not be controlled by cis-active elements within the first 2,400 bp of the promoter. Additional information may be required to restrict neuronal promoter expression to appropriate cell types. This regulatory information could reside elsewhere within the promoter, within introns, or may be provided by interactions between the two AADC promoters.
Aguanno
A
A
Roche Institute of Molecular Biology, Nutley, NJ 07110, USA.
Lee
M R
MR
Marden
C M
CM
Rattray
M
M
Gault
A
A
Albert
V R
VR
D000818:Animals; D001142:Aromatic-L-Amino-Acid Decarboxylases / Q000235:genetics*; D001483:Base Sequence; D018983:DNA Footprinting; D003850:Deoxyribonuclease I; D005796:Genes; D017403:In Situ Hybridization; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D009474:Neurons / Q000502:physiology*; D016716:PC12 Cells; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D012260:Ribonucleases
D015335:Molecular Probes; D012333:RNA, Messenger; D012260:Ribonucleases; D003850:Deoxyribonuclease I; D001142:Aromatic-L-Amino-Acid Decarboxylases