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  Characterization of the Functional Role of E-box Elements for the Transcriptional Activity of Rat Acetylcholine Receptor ε-Subunit and γ-Subunit Gene Promoters in Primary Muscle Cell Cultures

Dürr, I., Numberger, M., Berberich, C., & Witzemann, V. (1994). Characterization of the Functional Role of E-box Elements for the Transcriptional Activity of Rat Acetylcholine Receptor ε-Subunit and γ-Subunit Gene Promoters in Primary Muscle Cell Cultures. European Journal of Biochemistry, 224(2), 353-364. doi:10.1111/j.1432-1033.1994.00353.x.

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Dürr, Ingolf, Author
Numberger, Markus1, Author           
Berberich, Christoph1, 2, 3, Author           
Witzemann, Veit1, 2, 3, 4, Author           
Affiliations:
1Department of Cell Physiology, Max Planck Institute for Medical Research, Max Planck Society, ou_1497701              
2Molecular anatomy of the neuromuscular junction, Max Planck Institute for Medical Research, Max Planck Society, ou_1497727              
3Working Group Witzemann / Koenen, Max Planck Institute for Medical Research, Max Planck Society, ou_1497748              
4Department of Molecular Neurobiology, Max Planck Institute for Medical Research, Max Planck Society, ou_1497704              

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 Abstract: The expression of γ and ε subunits of the acetylcholine receptor from mammalian skeletal muscle is regulated independently during myogenic differentiation and innervation. Genomic DNA fragments containing 5′-flanking sequences of the ε-subunit and γ-subunit genes were characterised by a series of 5′ deletions fused to the chloramphenicol-acetyltransferase gene and transiently expressed by transfection of primary cultures of rat muscle cells and non-muscle cells. A 6.3-kb ε-subunit fragment can be reduced to yield a 270-bp fragment that confers 5–10-times higher expression levels in muscle cells compared to in non-muscle cells. The region composed of nucleotides –185 to –128 increases the transcriptional activity moderately while the 14-bp palindrome containing a single E box at nucleotides –88 to –83 may interact with the promoter but has no enhancer properties in muscle cells. From a 1.1-kb genomic fragment of the γ-subunit gene, 167 bp were sufficient for muscle-specific expression. Two promoter-proximal E-box elements enhance promoter activity in muscle and mediate transactivation by myogenic factors. Myogenin and myf5 were much more efficient than MRF4 or MyoD1 which exerted only little transactivation. Cotransfection experiments show that increased expression of Id in primary muscle cells inhibits chloramphenicol-acetyltransferase expression mediated by the γ-subunit gene promoter and support the view that myogenic factors play an important role in the transcriptional regulation of the γ-subunit gene.

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Language(s): eng - English
 Dates: 1994-04-272005-03-031994-09
 Publication Status: Issued
 Pages: 12
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 Rev. Type: Peer
 Identifiers: eDoc: 665152
DOI: 10.1111/j.1432-1033.1994.00353.x
Other: 6916
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Title: European Journal of Biochemistry
Source Genre: Journal
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Publ. Info: Berlin : Published by Springer-Verlag on behalf of the Federation of European Biochemical Societies
Pages: - Volume / Issue: 224 (2) Sequence Number: - Start / End Page: 353 - 364 Identifier: ISSN: 0014-2956
CoNE: https://pure.mpg.de/cone/journals/resource/111097776606040