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PMID: 8502560 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Negative regulatory elements upstream of a novel exon of the neuronal nicotinic acetylcholine receptor alpha 2 subunit gene.

Nucleic acids research ·Vol. 21 ·No. 9 ·1993-05-11 ·Pages 2185-92

Bessis A, Savatier N, Devillers-Thiéry A, Bejanin S, Changeux JP

Abstract

The expression of the nicotinic acetylcholine receptor alpha 2 subunit gene is highly restricted to the Spiriform lateralis nucleus of the Chick diencephalon. As a first step toward understanding the molecular mechanism underlying this regulation, we have investigated the structural and regulatory properties of the 5' sequence of this gene. A strategy based on the ligation of an oligonucleotide to the first strand of the cDNA (SLIC) followed by PCR amplification was used. A new exon was found approximately 3kb upstream from the first coding exon, and multiple transcription start sites of the gene were mapped. Analysis of the flanking region shows many consensus sequences for the binding of nuclear proteins, suggesting that the 1 kb flanking region contains at least a portion of the promoter of the gene. We have analysed the negative regulatory elements present within this region and found that a silencer region located between nucleotide -144 and +76 is active in fibroblasts as well as in neurons. This silencer is composed of six tandem repeat Oct-like motifs (CCCCATGCAAT), but does not bind any member of the Oct family. Moreover these motifs were found to act as a silencer only when they were tandemly repeated. When two, four or five motifs were deleted, the silencer activity of the motifs unexpectedly became an enhancer activity in all cells we have tested.

MeSH Terms
Animals Base Sequence Cell Line Chickens DNA DNA-Binding Proteins/metabolism Exons Introns Molecular Sequence Data Neurons/metabolism PC12 Cells Protein Biosynthesis Receptors, Nicotinic/genetics,metabolism Regulatory Sequences, Nucleic Acid Repetitive Sequences, Nucleic Acid
Chemicals
DNA-Binding Proteins Receptors, Nicotinic DNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Bessis A
UA CNRS D1284, Department des Biotechnologies, Institut Pasteur, Paris, France.
Savatier N
Devillers-Thiéry A
Bejanin S
Changeux J P
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/21.9.2185
Published
1993-05-11
Pages
2185-92
Language
English
Region
England
NLM ID
0411011
PMCID
PMC309483
Subset
IM
Databases
GENBANK
X69511
External Links
PubMed source DOI: 10.1093/nar/21.9.2185 PMC Full Text View journal details
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