A 5'-flanking region of the chicken acetylcholine receptor alpha-subunit gene confers tissue specificity and developmental control of expression in transfected cells.

Klarsfeld, André; Daubas, P; Bourachot, Brigitte; Changeux, J P

doi:10.1128/mcb.7.2.951

Cited by 91 publications

(60 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Rather, the CHX experiments reported here suggest that synthesis of an activator protein precedes the denervation-triggered receptor up-regulation. This agrees with the identification, in the promoter regions of the chick AChR ct- [16][17][18] Nuclease protection analysis was performed as described in Materials and Methods, with 30 p.g of total RNA and an excess of 10-fold or greater of radiolabeled riboprobe. Lane 1: intact ct-subunit probe; 2: molecular weight markers (¢~X174/HaelII); 3" CHX (at 0.2 mg/kg body weight) administered i.p.…”

Section: Chx Blocks Denervation-induced Rise In A-subunitsupporting

confidence: 79%

Protein synthesis is required for the denervation‐triggered activation of acetylcholine receptor genes

1990

View full text Add to dashboard Cite

The effect of cycloheximide (CHX) on denervation-induced acetylcholine receptor (AChR) expression was investigated in chickens one day after nerve section, using probe excess solution hybridization to quantitate AChR ct-subunit gene transcript levels and run-on analysis to measure subunit gene activity. The increase in ct-subunit transcripts that normally follows denervation was prevented when drug treatment was commenced 2 h before or after denervation but was not blocked when CHX administration was begun 6 h after the operation. Drug-induced reduction of transcript levels results from decreased activity of genes coding for the ~t-, 6-, and y-subunits; in contrast, the transcription rates of several non-receptor genes are not affected by CHX. The results suggest that the de novo synthesis of a transcriptional activator is required as a mediating event in the signalling pathway linking the plasma membrane and AChR gene expression.Transcriptional activator; Subunit mRNA metabolism; Cycloheximide; Chick muscle

show abstract

Section: Chx Blocks Denervation-induced Rise In A-subunitsupporting

confidence: 79%

Protein synthesis is required for the denervation‐triggered activation of acetylcholine receptor genes

1990

View full text Add to dashboard Cite

show abstract

“…12). The region between -1242 and -1232 shares 80% homology with a 10-bp element upstream of the chicken ACh receptor asubunit gene (29). An 8-bp segment of this sequence from the ACh receptor upstream region is repeated three times in the MCK proximal region, where it overlaps with three 17-bp imperfect repeats that were identified previously (28) to be associated with other muscle-specific genes (see Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The identification of such homology suggests a strong evolutionary constraint on these sequences and has led to speculation concerning their potential involvement in muscle-specific regulation. Transfection studies of muscle cells in tissue culture also have demonstrated sequences adjacent to or within the genes encoding skeletal cx actin (3,18,35,45), cardiac a actin (23,37,38), the ACh receptor a subunit (29), MCK (28), Tn-I (30,31), MHC (6), and MLC type 2 (MLC-2) (43) that are sufficient to direct differentiation-specific expression. Sequences upstream from a subset of these genes also have been shown to direct appropriate expression in transgenic mice (60,61), Xenopus oocytes (63), and Drosophila melanogaster (15).…”

mentioning

confidence: 99%

Identification of upstream and intragenic regulatory elements that confer cell-type-restricted and differentiation-specific expression on the muscle creatine kinase gene.

Sternberg¹,

Spizz²,

Perry³

et al. 1988

Mol. Cell. Biol.

250

207

View full text Add to dashboard Cite

Terminal differentiation of skeletal myoblasts is accompanied by induction of a series of tissue-specific gene products, which includes the muscle isoenzyme of creatine kinase (MCK). To begin to define the sequences and signals involved in MCK regulation in developing muscle cells, the mouse MCK gene has been isolated. Sequence analysis of 4,147 bases of DNA surrounding the transcription initiation site revealed several interesting structural features, some of which are common to other muscle-specific genes and to cellular and viral enhancers. To test for sequences required for regulated expression, a region upstream of the MCK gene from -4800 to +1 base pairs, relative to the transcription initiation site, was linked to the coding sequences of the bacterial chloramphenicol acetyltransferase (CAT) gene. Introduction of this MCK-CAT fusion gene into C2 muscle cells resulted in high-level expression of CAT activity in differentiated myotubes and no detectable expression in proliferating undifferentiated myoblasts or in nonmyogenic cell lines. Deletion mutagenesis of sequences between -4800 and the transcription start site showed that the region between -1351 and -1050 was sufficient to confer cell type-specific and developmentally regulated expression on the MCK promoter. This upstream regulatory element functioned independently of position, orientation, or distance from the promoter and therefore exhibited the properties of a classical enhancer. This upstream enhancer also was able to confer muscle-specific regulation on the simian virus 40 promoter, although it exhibited a 3- to 5-fold preference for its own promoter. In contrast to the cell type- and differentiation-specific expression of the upstream enhancer, the MCK promoter was able to function in myoblasts and myotubes and in nonmyogenic cell lines when combined with the simian virus 40 enhancer. An additional positive regulatory element was identified within the first intron of the MCK gene. Like the upstream enhancer, this intragenic element functioned independently of position, orientation, and distance with respect to the MCK promoter and was active in differentiated myotubes but not in myoblasts. These results demonstrate that expression of the MCK gene in developing muscle cells is controlled by complex interactions among multiple upstream and intragenic regulatory elements that are functional only in the appropriate cellular context.

show abstract

“…2A). ␣1KK encompasses nucleotides 151-334 of the chick ␣1 nAChR promoter (19,20) (GenBank TM accession number M15307), flanked by KpnI restriction sites that were used for subcloning into pCAT00. The ␣1/␤3 hybrid promoter was obtained by ligation of the 5Ј-end of ␣1KK with the 3Ј-end of ␤3RS at their shared PvuII restriction site (Fig.…”

Section: Methodsmentioning

confidence: 99%

Functional Properties of the Neuronal Nicotinic Acetylcholine Receptor β3 Promoter in the Developing Central Nervous System

Roztocil

Matter-Sadzinski

Gomez

et al. 1998

Journal of Biological Chemistry

View full text Add to dashboard Cite

Within the chick central nervous system, expression of the ␤3 nicotinic acetylcholine receptor gene is restricted to a subset of retinal neurons, the majority of which are ganglion cells. Transient transfection in retinal neurons and in neural and non-neural cells from other regions of the chick embryo allowed the identification of the cis-regulatory domain of the ␤3 gene. Within this domain, a 75-base pair fragment located immediately upstream of the transcription start site suffices to reproduce the neuron-specific expression pattern of ␤3. This fragment encompasses an E-box and a CAAT box, both of which are shown to be key positive regulatory elements of the ␤3 promoter. Co-transfection experiments into retinal, telencephalic, and tectal neurons with plasmid reporters of ␤3 promoter activity and a number of vectors expressing different neuronal (ASH-1, NeuroM, NeuroD, CTF-4) and non-neuronal (MyoD) basic helix-loop-helix transcription factors indicate that the cis-regulatory domain of ␤3 has the remarkable property of discriminating accurately between related members of the basic helix-loop-helix protein family. The sequence located immediately 3 of the E-box participates in this selection, and the E-box acts in concert with the nearby CAAT box.In vertebrates, both negative and positive regulations play an important role in determining neuronal gene expression. Negative regulation (reviewed in Refs. 1 and 2) is best exemplified by REST/NRSF, a factor that represses transcription of the SCG-10 and type II Na ϩ channel genes in non-neuronal cell-types, whereas most neurons lack REST/NRSF and thus express these two genes (3, 4). Conversely, the Olf-1 transcription factor positively regulates several genes (e.g. the olfactory neuron-specific G protein, the type III adenylyl cyclase) specifically expressed in olfactory neurons (5). Vertebrate homologs of the basic helix-loop-helix (bHLH) 1 factors involved in Drosophila neurogenesis (6) act as positive or negative regulators in the acquisition of neuronal identity. For instance, the atonaland achaete-scute-related activators are transiently expressed in parts of the central and peripheral nervous system during early development, and their null mutation or ectopic expression profoundly influences neurogenesis (reviewed by Lee et al. (7)). However, the direct regulation by bHLH proteins of genes that define neuronal identity has never been documented.Several neuronal nicotinic acetylcholine receptor (nAChR) genes are expressed early in neural development (8 -12), and, since they encode transmembrane sensors capable of fluxing Ca 2ϩ and other cations upon stimulation (reviewed in Ref. 13), an understanding of their regulation should help explain how the genetic program puts together the mechanisms needed for epigenetic environmental cues to participate in development. This is illustrated in a recent report by Shatz and associates (14) showing that, in the developing retina, cholinergic synaptic transmission between newly generated amacrine and ganglion cells is respo...

show abstract

A 5'-flanking region of the chicken acetylcholine receptor alpha-subunit gene confers tissue specificity and developmental control of expression in transfected cells.

Cited by 91 publications

References 33 publications

Protein synthesis is required for the denervation‐triggered activation of acetylcholine receptor genes

Protein synthesis is required for the denervation‐triggered activation of acetylcholine receptor genes

Identification of upstream and intragenic regulatory elements that confer cell-type-restricted and differentiation-specific expression on the muscle creatine kinase gene.

Functional Properties of the Neuronal Nicotinic Acetylcholine Receptor β3 Promoter in the Developing Central Nervous System

Contact Info

Product

Resources

About