Gene Structure of the Rat Kainate Receptor Subunit KA2 and Characterization of an Intronic Negative Regulatory Region

Huang, Fei; Gallo, Vittorio

doi:10.1074/jbc.272.13.8618

Cited by 16 publications

(31 citation statements)

References 38 publications

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“…We have also identified an intronic element of this gene, which displays functional features of a silencer (31). More recently, we described the binding of nuclear orphan receptor proteins to this sequence to down-regulate GRIK5 transcription (35).…”

mentioning

confidence: 93%

“…Cell Cultures-CG-4, HeLa, and NIH3T3 cells were cultured as previously described (31,37). Cortical astrocytes were prepared as previously described (38,39).…”

Section: Methodsmentioning

confidence: 99%

“…The isolation and characterization of GRIK5 genomic clones from a rat Sprague-Dawley genomic library (DASH II; Stratagene, La Jolla, CA) have been described previously (31). The nucleotide sequence reported in this paper has been previously submitted to the GenBank™/EBI Data Bank with accession number U81010.…”

Section: Grik5 Reporter Gene Constructs and Transient Transfection Asmentioning

confidence: 99%

“…The kainate receptor subunit KA2 is encoded by the GRIK5 gene (30,31). KA2 transcripts can be detected in the rat as early as embryonic day 10 (E10) in the neural tube (32) and at E12 in the cortical plate (9).…”

mentioning

confidence: 99%

“…To gain an understanding of how kainate receptor subunit expression is regulated, we have previously cloned the GRIK5 gene and characterized its structure (31). We have also identified an intronic element of this gene, which displays functional features of a silencer (31).…”

mentioning

confidence: 99%

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Characterization of the Rat GRIK5 Kainate Receptor Subunit Gene Promoter and Its Intragenic Regions Involved in Neural Cell Specificity

Chew

Yuan

Scherer

et al. 2001

Journal of Biological Chemistry

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The GRIK5 (glutamate receptor ionotropic kainate-5) gene encodes the kainate-preferring glutamate receptor subunit KA2. The GRIK5 promoter is TATA-less and GCrich, with multiple consensus initiator sequences. Transgenic mouse lines carrying 4 kilobases of the GRIK5 5-flanking sequence showed lacZ reporter expression predominantly in the nervous system. Reporter assays in central glial (CG-4) and non-neural cells indicated that a 1200-base pair (bp) 5-flanking region could sustain neural cell-specific promoter activity. Transcriptional activity was associated with the formation of a transcription factor IID-containing complex on an initiator sequence located 1100 bp upstream of the first intron. In transfection studies, deletion of exonic sequences downstream of the promoter resulted in reporter gene activity that was no longer neural cell-specific. When placed downstream of the GRIK5 promoter, a 77-bp sequence from the deleted fragment completely silenced reporter expression in NIH3T3 fibroblasts while attenuating activity in CG-4 cells. Analysis of the 77-bp sequence revealed a functional SP1-binding site and a sequence resembling a neuron-restrictive silencer element. The latter sequence, however, did not display cell-specific binding of REST-like proteins. Our studies thus provide evidence for intragenic control of GRIK5 promoter activity and suggest that elements contributing to tissue-specific expression are contained within the first exon.Three structurally related mammalian glutamate receptor (GluR) 1 gene families encode N-methyl-D-aspartate, ␣-amino-3-hydroxymethylisoxazole-4-propionic acid, and kainate receptor subunits (1-3). Two kainate receptor gene subfamilies encode the subunits GluR5-7 and KA1/2, which bind kainate and domoate with high affinity (2-4). When expressed in Xenopus oocytes or heterologous mammalian cells, only GluR5-7 form functional homomeric channels (5-7). KA1 and KA2 are inactive as homomeric ion channels, but associate with members of the GluR5-7 family to form functional heteromeric kainatepreferring receptors with biophysical properties and agonist selectivity distinct from those of homomeric GluR5-7 channels (8 -10).In situ hybridization and immunohistochemical studies have demonstrated that, in different brain regions, specific combinations of members of the KA1/2 and GluR5-7 gene subfamilies are coexpressed (11-13). This indicates that, similar to ␣-amino-3-hydroxymethylisoxazole-4-propionic acid and Nmethyl-D-aspartate receptors, kainate receptor subunits assemble to form diverse subtypes of heteromeric channels. For example, co-immunoprecipitation experiments with antiGluR6 and anti-KA2 antibodies have shown that these subunit proteins associate to form receptor channel complexes in brain cell membranes (14).These molecular studies are consistent with the finding that functional homomeric and hetero-oligomeric kainate-preferring receptors are expressed in cultured neurons and glia (15)(16)(17). Despite the widespread expression of kainate receptor subunits in the mammalia...

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mentioning

confidence: 93%

“…Cell Cultures-CG-4, HeLa, and NIH3T3 cells were cultured as previously described (31,37). Cortical astrocytes were prepared as previously described (38,39).…”

Section: Methodsmentioning

confidence: 99%

Section: Grik5 Reporter Gene Constructs and Transient Transfection Asmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Characterization of the Rat GRIK5 Kainate Receptor Subunit Gene Promoter and Its Intragenic Regions Involved in Neural Cell Specificity

Chew

Yuan

Scherer

et al. 2001

Journal of Biological Chemistry

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Expression and regulation of kainate and AMPA receptors in the rat neural tube

Scherer

Gallo

1998

J. Neurosci. Res.

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We analyzed the expression and regulation of glutamate receptor subunits in the rat neural tube (10 day embryos) and in cell cultures derived from this tissue. In the cultures, all cells were stained with antibodies against the neural progenitor marker nestin. More than 50% of the cells were also stained by the monoclonal antibodies LB1 or A2B5, which bind to neuronal and glial progenitors. Approximately 6% of the cells were stained with antibodies for the low affinity NGF receptor, a neural crest cell marker. A small percentage of cells differentiated to neurons or astrocytes, as determined by staining with anti-neurofilament and anti-GFAP antibodies, respectively. RT-PCR analysis of neural tube tissue and culture mRNAs demonstrated that the AMPA receptor subunits GluR3 and 4 and the kainate receptor subunits GluR6, 7, KA1 and KA2 were detectable at E10. The kainate receptor subunits GluR6 and KA2 were upregulated by culture conditions which stimulated cell differentiation, as determined by concomitant downregulation of nestin mRNA. Both in neural tube tissue and in cultured cells, GluR6 was 100% unedited. Finally, both GluR6 and KA2 proteins could be detected in subpopulations of neural progenitors and differentiated neurons. Our data indicate that kainate receptor genes are expressed in undifferentiated progenitor cells of the neural tube at E10, and are upregulated during neural cell differentiation.

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Refined linkage to the RDP/DYT12 locus on 19q13.2 and evaluation of GRIK5 as a candidate gene

Kamm¹,

Leung²,

Joseph³

et al. 2004

Movement Disorders

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Gene Structure of the Rat Kainate Receptor Subunit KA2 and Characterization of an Intronic Negative Regulatory Region

Cited by 16 publications

References 38 publications

Characterization of the Rat GRIK5 Kainate Receptor Subunit Gene Promoter and Its Intragenic Regions Involved in Neural Cell Specificity

Characterization of the Rat GRIK5 Kainate Receptor Subunit Gene Promoter and Its Intragenic Regions Involved in Neural Cell Specificity

Expression and regulation of kainate and AMPA receptors in the rat neural tube

Refined linkage to the RDP/DYT12 locus on 19q13.2 and evaluation of GRIK5 as a candidate gene

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