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Multiple promoter elements required for leukemia inhibitory factor‐stimulated M 2 muscarinic acetylcholine receptor promoter activity
Author(s) -
Laszlo George S.,
Rosoff Marc L.,
Amieux Paul S.,
Nathanson Neil M.
Publication year - 2006
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.1471-4159.2006.03976.x
Subject(s) - muscarinic acetylcholine receptor , inhibitory postsynaptic potential , leukemia inhibitory factor , muscarinic acetylcholine receptor m1 , acetylcholine , muscarinic acetylcholine receptor m3 , muscarinic acetylcholine receptor m5 , acetylcholine receptor , muscarinic acetylcholine receptor m4 , chemistry , leukemia inhibitory factor receptor , neuroscience , biology , receptor , pharmacology , biochemistry , gene , embryonic stem cell
Treatment of neuronal cells with leukemia inhibitory factor (LIF) results in increased M 2 muscarinic acetylcholine receptor promoter activity. We demonstrate here that multiple promoter elements mediate LIF stimulation of M 2 gene transcription. We identify a LIF inducible element (LIE) in the M 2 promoter with high homology to a cytokine‐inducible ACTG‐containing sequence in the vasoactive intestinal peptide promoter. Mutagenesis of both a STAT (signal transducers and activators of transcription) element and the LIE in the M 2 promoter is required to attenuate stimulation of M 2 promoter activity by LIF completely. Mobility shift assays indicate that a LIF‐stimulated complex binds to a 70 base pair M 2 promoter fragment. Furthermore, a STAT element within this fragment can bind to LIF‐stimulated nuclear STAT1 homodimers in vitro . Mutagenesis experiments show that cytokine‐stimulated activation of M 2 promoter activity requires tyrosine residues on glycoprotein 130 (gp130) that are also required for both STAT1 and STAT3 activation. Dominant negative STAT1 or STAT3 can block LIF‐stimulated M 2 promoter activity. Real‐time RT‐PCR analysis indicates that LIF‐stimulated induction of M 2 mRNA is partially dependent on protein synthesis. These results show that regulation of M 2 gene transcription in neuronal cells by LIF occurs through a complex novel mechanism that is dependent on LIE, STAT and de novo protein synthesis.