A 102 base pair sequence of the nicotinic acetylcholine receptor delta-subunit gene confers regulation by muscle electrical activity
| Autor: | Kenneth G. Chahine, Daniel Goldman, Wade Walke |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
Expression vector
Base Sequence Transcription Genetic Myogenesis Muscles Molecular Sequence Data Structural gene Promoter Receptors Nicotinic Biology Molecular biology Acetylcholine Electric Stimulation Rats Electrophysiology Nicotinic acetylcholine receptor Gene Expression Regulation Gene expression Animals Luciferase Molecular Biology Cells Cultured Developmental Biology Acetylcholine receptor |
| Zdroj: | Development. 115:213-219 |
| ISSN: | 1477-9129 0950-1991 |
| DOI: | 10.1242/dev.115.1.213 |
| Popis: | Muscle electrical activity suppresses expression of the embryonic-type (α, β, γ and δ) nicotinic acetylcholine receptor (nAChR) genes. The molecular mechanism by which electrical activity regulates these genes is not known. One approach to this problem is to identify regions of the nAChR genes that mediate electrical regulation. Here we report results from such a study of the nAChR δ-subunit gene. We cloned the rat δ-subunit promoter region and created an expression vector in which this DNA controlled the expression of a down-stream luciferase structural gene. The effect that muscle electrical activity had on the expression from this promoter was assayed by introducing this expression vector into electrically stimulated and tetrodotoxin (TTX)-treated rat primary myotubes, and assaying for luciferase activity. These myotubes, when stimulated with extracellular electrodes, suppressed endogenous embryonic-type nAChR gene expression compared to those treated with TTX. Transfection of these cells with δ-promoter-luciferase expression vectors resulted in the δ-promoter conferring electrical regulation on luciferase expression. Additional experiments using deletions from the 5’ end of theδ-promoter region have identified an element between −677 and −550 bp that suppressed δ-promoter activity and a minimal 102 bp sequence that promotes and regulates luciferase expression in response to muscle electrical activity. This latter sequence also contains all the necessary elements to confer tissue and developmental stage-specific expression. |
| Databáze: | OpenAIRE |
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