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  • The bromodomain-containing gene BRD2 is regulated at transcription, splicing, and translation levels.

The bromodomain-containing gene BRD2 is regulated at transcription, splicing, and translation levels.

Journal of cellular biochemistry (2011-05-25)
Enyuan Shang, Qingping Cui, Xiangyuan Wang, Cheryl Beseler, David A Greenberg, Debra J Wolgemuth
ABSTRACT

The human BRD2 gene has been linked and associated with a form of common epilepsy and electroencephalographic abnormalities. Disruption of Brd2 in the mouse revealed that it is essential for embryonic neural development and that viable Brd2(+/-) heterozygotes show both decreased GABAergic neuron counts and increased susceptibility to seizures. To understand the molecular mechanisms by which mis-expression of BRD2 might contribute to epilepsy, we examined its regulation at multiple levels. We discovered that BRD2 expresses distinct tissue-specific transcripts that originate from different promoters and have strikingly different lengths of 5' untranslated regions (5'UTR). We also experimentally confirmed the presence of a highly conserved, alternatively spliced exon, inclusion of which would result in a premature termination of translation. Downstream of this alternative exon is a polymorphic microsatellite (GT-repeats). Manipulation of the number of the GT-repeats revealed that the length of the GT-repeats affects the ratio of the two alternative splicing products. In vitro translation and expression in cultured cells revealed that among the four different mRNAs (long and short 5'UTR combined with regular and alternative splicing), only the regularly spliced mRNA with the short 5'UTR yields full-length protein. In situ hybridization and immunohistochemical studies showed that although Brd2 mRNA is expressed in both the hippocampus and cerebellum, Brd2 protein only can be detected in the cerebellar Purkinje cells and not in hippocampal cells. These multiple levels of regulation would likely affect the production of functional BRD2 protein during neural development and hence, its role in the etiology of seizure susceptibility.

MATERIALS
Product Number
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Product Description

Sigma-Aldrich
Monoclonal ANTI-FLAG® M2 antibody produced in mouse, clone M2, purified immunoglobulin (Purified IgG1 subclass), buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide)
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BRD2 (339-459) His tag human, recombinant, expressed in E. coli, ≥90% (SDS-PAGE)
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BRD2 (65-187) GST tag human, recombinant, expressed in E. coli, ≥81% (SDS-PAGE)
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BRD2 (339-459) GST tag human, recombinant, expressed in E. coli, ≥86% (SDS-PAGE)
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BRD2, BD1, BD2 (65-459) GST tag human, recombinant, expressed in E. coli, ≥80% (SDS-PAGE)
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BRD2 (65-187) His tag human, recombinant, expressed in E. coli, ≥90% (SDS-PAGE)