Analysis of the alternative promoters that regulate tissue-specific expression of human aromatic L-amino acid decarboxylase.
Previously we identified two alternative first exons (exon N1 and exon L1) coding for 5' untranslated regions of human aromatic L-amino acid decarboxylase (AADC) and found that their alternative usage produced two types of mRNAs in a tissue-specific manner. To determine the cis-acting element regulating the tissue-specific expression of human AADC, we produced three kinds of transgenic mice harboring 5' flanking regions of the human AADC gene fused to the bacterial chloramphenicol acetyltransferase (CAT) gene. The transgene termed ACA contained -7.0 kb to -30 bp in exon N1, including the entire exon L1; ACN contained -3.6 kb to -30 bp in exon N1; and ACL contained -2.8 kb to -42 bp in exon L1. The ACA transgenic mice expressed CAT at extremely high levels in peripheral nonneuronal tissues, such as pancreas, liver, kidney, small intestine, and colon, that contained endogenous high AADC activity, whereas CAT immunoreactivity was not detected in either catecholaminergic or serotonergic neurons in the CNS. Thus, it was suggested that the ACA transgene contained the major part of cis-regulatory elements for the expression of AADC in peripheral nonneuronal tissues. On the other hand, the ACN transgenic mice moderately expressed CAT in various tissues except for the lung and liver, and the ACL transgenic mice showed moderate CAT expression only in the kidney.
Sumi-Ichinose
C
C
Department of Pharmacology, School of Medicine, Fujita Health University, Aichi, Japan.
Hasegawa
S
S
Ichinose
H
H
Sawada
H
H
Kobayashi
K
K
Sakai
M
M
Fujii
T
T
Nomura
H
H
Nomura
T
T
Nagatsu
I
I
D000818:Animals; D001142:Aromatic-L-Amino-Acid Decarboxylases / Q000235:genetics*; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008822:Mice, Transgenic; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic
D015335:Molecular Probes; D015500:Chloramphenicol O-Acetyltransferase; D001142:Aromatic-L-Amino-Acid Decarboxylases