DNA-bound transcription factor complexes analysed by mass-spectrometry: binding of novel proteins to the human c-fos SRE and related sequences

Drewett, Victoria; Molina, Henrik; Millar, Alan; Muller, Silke; Hesler, Friedrich von; Shaw, Peter E.

doi:10.1093/nar/29.2.479

Cited by 39 publications

(27 citation statements)

References 34 publications

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“…DNA Pulldown Assay-DNA pulldown assay was performed as described previously (40). Oligonucleotide duplexes corresponding to the sequence of the CRE site in the ␣1 promoter and its flanking region were covalently linked to a biotin moiety at their 5Ј ends.…”

Section: Methodsmentioning

confidence: 99%

Surface Expression of GABAA Receptors Is Transcriptionally Controlled by the Interplay of cAMP-response Element-binding Protein and Its Binding Partner Inducible cAMP Early Repressor

Lund²,

Gravielle

et al. 2008

Journal of Biological Chemistry

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The regulated expression of type A ␥-aminobutyric acid (GABA) receptor (GABA A R) subunit genes plays a critical role in neuronal maturation and synaptogenesis. It is also associated with a variety of neurological diseases. Changes in GABA A receptor ␣1 subunit gene (GABRA1) expression have been reported in animal models of epilepsy, alcohol abuse, withdrawal, and stress. Understanding the genetic mechanism behind such changes in ␣ subunit expression will lead to a better understanding of the role that signal transduction plays in control over GABA A R function and brings with it the promise of providing new therapeutic tools for the prevention or cure of a variety of neurological disorders. Here we show that activation of protein kinase C increases ␣1 subunit levels via phosphorylation of CREB (pCREB) that is bound to the GABRA1 promoter (GABRA1p). In contrast, activation of protein kinase A decreases levels of ␣1 even in the presence of pCREB. Decrease of ␣1 is dependent upon the inducible cAMP early repressor (ICER) as directly demonstrated by ICER-induced down-regulation of endogenous ␣1-containing GABA A Rs at the cell surface of cortical neurons. Taken together with the fact that there are less ␣1␥2-containing GABA A Rs in neurons after protein kinase A stimulation and that activation of endogenous dopamine receptors down-regulates ␣1 subunit mRNA levels subsequent to induction of ICER, our studies identify a transcriptional mechanism for regulating the cell surface expression of ␣1-containing GABA A Rs that is dependent upon the formation of CREB heterodimers. ␥-Aminobutyric acid (GABA)4 type A receptors (GABA A Rs) are ligand-gated chloride ion channels that mediate the majority of fast synaptic inhibition in the mammalian brain (1). They are pentameric in structure and are composed of multiple subunit isoforms coming from eight distinct classes: ␣, ␤, ␥, ␦, , ⑀, , and . Diversity in receptor subtypes is controlled by the regulated expression of 19 different subunit genes and the alternative splicing of individual subunit transcripts (2-4). Most importantly, differential receptor subunit composition produces functionally and pharmacologically distinct GABA A Rs at certain times during development and in certain regions of the brain (5-8).The transcription of different GABA A R subunit genes (GABRs) is likely to involve a complex system of regulatory controls that remain to be identified. Several lines of evidence suggest that ␣1 subunit expression is activity-dependent (9 -11). Treatment with N-methyl-D-aspartate, a selective activator of an important class of excitatory ligand-gated ion channels, stimulates ␣1 subunit expression in cultured cerebellar granule cells (12, 13). In contrast, chronic treatment of cortical neurons with GABA decreases levels of GABA A R ␣1 subunit mRNAs (14 -16), which is dependent on voltage-gated calcium channel activity and most likely an alteration in transcription (15). In addition, prolonged benzodiazepine (BZ) treatment decreases ␣1 mRNA levels in the rat hippocampal CA...

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Section: Methodsmentioning

confidence: 99%

Surface Expression of GABAA Receptors Is Transcriptionally Controlled by the Interplay of cAMP-response Element-binding Protein and Its Binding Partner Inducible cAMP Early Repressor

Lund²,

Gravielle

et al. 2008

Journal of Biological Chemistry

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“…The resulting biotinylated IL-1␤131 fragments are designated IL-1␤131 wt, m45 (mutated fragment in the PU.1 Ϫ45 site), m90 (mutated fragment in the C/EBP Ϫ90 site), and dm (doubly mutated at both the Ϫ45 and Ϫ90 sites). Generation of a biotinylated TPA response element (TRE) was obtained by Klenow fill-in of the 5Ј overhang created after annealing of the sense AATTCTTCCGGCTGACTC ATCAAGC (AP-1 site is underlined) and antisense GCTTGATGAGTCAGCC GGAAG oligonucleotides as described previously (16). Equal molar quantities of biotinylated DNA templates were than immobilized on magnetic resins conjugated with streptavidin according to the manufacturer's instructions (IL-1␤131 short and TRE; final concentration, 0.06 pmol/l).…”

mentioning

confidence: 99%

c-Jun Homodimers Can Function as a Context-Specific Coactivator

Grondin

Lefrançois

Tremblay

et al. 2007

Molecular and Cellular Biology

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“…It would be of particular interest to identify other factors that may interact with Miz-1 under these conditions; alternatively, other Inr-binding proteins could facilitate such gene stimulatory events. Given the latest developments in proteomics and the identification of DNA-protein complexes (44), it should be ultimately feasible to elucidate factors bound at particular core promoters during the various stages of macrophage activation. Similarly, techniques such as RNA interference may tell us which Inr-binding factors are important for Nramp1 promoter function under a variety of conditions.…”

Section: Fig 6 Responses Of the Nramp1mentioning

confidence: 99%

Characterization of the Murine Nramp1 Promoter

Bowen

Lapham

Phillips

et al. 2003

Journal of Biological Chemistry

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Murine Nramp1 encodes a divalent cation transporter that is expressed in late endosomes/lysosomes of macrophages, and the transported cations facilitate intracellular pathogen growth control. The Nramp1 promoter is TATA box-deficient, has two initiator elements, and is repressed by c-Myc, in accordance with the notion that genes that deplete the iron content of the cell cytosol antagonize cell growth. Repression via c-Myc occurs at the initiator elements, whereas a c-Myc-interacting protein (Miz-1) stimulates transcription. Here we demonstrate that a non-canonical E box (CAACTG) inhibits basal promoter activity and activation by Miz-1. A consensus Sp1-binding site or GC box is also necessary for Miz-1-dependent transactivation, but not repression. Repression occurs by c-Myc competing with p300/CBP for binding Miz-1. Our results show that an Sp1 site mutant inhibits coactivation by p300 and that the murine Nramp1 promoter is preferentially expressed within macrophages (relative to a ␤-actin control) compared with non-macrophage cells. The effect of the Sp1 site mutation on promoter function shows cell-type specificity: stimulation in COS-1 and inhibition in RAW264.7 cells. Miz-1-directed RNA interference confirms a stimulatory role for Miz-1 in Nramp1 promoter function. c-Myc, Miz-1, and Sp1 were identified as binding to the Nramp1 core promoter in control cells and following acute stimulation with interferon-␥ and lipopolysaccharide. These results provide a description of sites that modulate the activity of the initiator-binding protein Miz-1 and indicate a stimulatory role for GC box-binding factors in macrophages and a inhibitory role for E box elements in proliferating cells. The murine Nramp1/Slc11a11 locus encodes a proton-coupled divalent cation transporter, expressed in late endosomes/ lysosomes of macrophages, that constitutes a component of the innate immune response to combat intracellular pathogens (1, 2). Nramp1 is biallelic in mouse; one allele, G169, restricts pathogen growth (termed resistant), whereas allele D169 (susceptible/functionally null) is permissive for pathogen growth (3). We have proposed that Nramp1 plays an important role in iron transport associated with homeostasis and pathology, in addition to its established role in infection (4), although the precise function of Nramp1 in transporting erythrocyte-acquired iron has still to be clarified, particularly with regard to the interrelationship with heme oxygenases. There is continuing controversy regarding the mechanism of the antibacterial activity of Nramp1. Some data support an iron starvation activity for Nramp1 (5, 6), whereas others indicate Fenton chemistry-mediated killing within the lumen of the phagosome, catalyzed by Nramp1-transported iron (7). Our data are consistent with Nramp1 depleting the iron content of the macrophage cytosol (8) and with the divalent cation proton antiport transport mechanism for Nramp1 proposed by Blackwell and co-workers (9).Allelic Nramp1 is associated with many pleiotropic effects within the macro...

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DNA-bound transcription factor complexes analysed by mass-spectrometry: binding of novel proteins to the human c-fos SRE and related sequences

Cited by 39 publications

References 34 publications

Surface Expression of GABAA Receptors Is Transcriptionally Controlled by the Interplay of cAMP-response Element-binding Protein and Its Binding Partner Inducible cAMP Early Repressor

Surface Expression of GABAA Receptors Is Transcriptionally Controlled by the Interplay of cAMP-response Element-binding Protein and Its Binding Partner Inducible cAMP Early Repressor

c-Jun Homodimers Can Function as a Context-Specific Coactivator

Characterization of the Murine Nramp1 Promoter

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