Promoter elements of the rat choline acetyltransferase gene allowing nerve growth factor inducibility in transfected primary cultured cells.
Choline acetyltransferase, the enzyme responsible for the synthesis of acetylcholine, provides a convenient index for cholinergic neurons. Using a previously identified rat cDNA clone, we have isolated several corresponding genomic clones and have characterized a 1,902-bp fragment that contains part of the first noncoding exon as well as promoter sequences. The promoter activity of this fragment was tested, taking advantage of the recently developed lipopolyamine-mediated DNA transfer method, which allows transfection of primary neurons. The 1,902-bp sequence drives the expression of the bacterial chloramphenicol acetyltransferase (CAT) reporter gene in a culture of dissociated cells prepared from the septal area of fetal (embryonic day 17) rats, a structure rich in cholinergic neurons. Moreover, addition of nerve growth factor to the culture increases CAT expression by approximately 56-fold, indicating that our DNA fragment contains sequences required for NGF induction. In addition, it contains consensus sequences for various transcription factors, including those of the basic helix-loop-helix family. Finally, experiments to characterize the transcription start site are presented.
Bejanin
S
S
Departement de Génétique Moléculaire, CNRS, Gif-sur-Yvette, France.
Habert
E
E
Berrard
S
S
Edwards
J B
JB
Loeffler
J P
JP
Mallet
J
J
D000818:Animals; D001483:Base Sequence; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D002795:Choline O-Acetyltransferase / Q000235:genetics*; D002874:Chromosome Mapping; D005796:Genes; D016678:Genome; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012688:Septum Pellucidum / Q000166:cytology / Q000196:embryology; D014162:Transfection
D009414:Nerve Growth Factors; D002795:Choline O-Acetyltransferase