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Multiple promoters drive tissue‐specific expression of the human M 2 muscarinic acetylcholine receptor gene
Author(s) -
Krejci Alena,
Bruce Alexander W.,
Dolezal Vladimir,
Tucek Stanislav,
Buckley Noel J.
Publication year - 2004
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.2004.02694.x
Subject(s) - muscarinic acetylcholine receptor , promoter , acetylcholine , acetylcholine receptor , muscarinic acetylcholine receptor m2 , muscarinic acetylcholine receptor m4 , muscarinic acetylcholine receptor m5 , gene expression , muscarinic acetylcholine receptor m3 , gene , muscarinic acetylcholine receptor m1 , microbiology and biotechnology , biology , neuroscience , receptor , chemistry , genetics , pharmacology
Despite the wealth of information on the functional and pharmacological properties of the M 2 muscarinic receptor, we know relatively little of structure and regulation of the M 2 receptor gene. Here, we describe the organisation of the human M 2 gene and its promoters. Four exons are present in the 5′ untranslated region of the human M 2 mRNA distributed over 146 kb on chromosome 7 which produce eight different splice variants in the IMR‐32 neuroblastoma cell line. The unexpectedly large size of this gene indicates that transcription initiates much further upstream of the coding region than earlier studies had indicated. We present evidence that there are three distinct human M 2 promoters. Analysis of endogenous transcripts revealed that promoter 2 is preferentially used in neuroblastoma cells, whereas promoter 1 in cardiac cells. All promoters are highly conserved across human, mouse, rat and pig. They contain multiple start sites and none possess a TATA‐box. In addition, we describe another M 2 promoter that is specific for rat. We show that GATA‐4 transcription factor binds to two sites within the regulatory regions of the M 2 gene using reporter gene assays, electromobility shift assays and mutational analysis.