Division of Statistical Genetics, Department of Biostatistics, Mailman School of Public Health, Columbia University Medical Center, New York, New York 10032, USA.
J Cell Biochem. 2011 Oct;112(10):2784-93. doi: 10.1002/jcb.23192.
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.
人类 BRD2 基因与一种常见的癫痫和脑电图异常有关联。在小鼠中破坏 Brd2 发现,它对胚胎神经发育至关重要,而存活的 Brd2(+/-)杂合子表现出 GABA 能神经元计数减少和癫痫易感性增加。为了了解 BRD2 表达异常可能导致癫痫的分子机制,我们在多个层面检查了它的调节。我们发现 BRD2 表达出独特的组织特异性转录本,这些转录本来自不同的启动子,并且 5'非翻译区(5'UTR)的长度有明显差异。我们还通过实验证实了存在一个高度保守的、可变剪接的外显子,包含该外显子会导致翻译过早终止。在这个替代外显子的下游是一个多态性微卫星(GT-重复)。对 GT-重复数的操纵表明,GT-重复的长度会影响两种选择性剪接产物的比例。体外翻译和在培养细胞中的表达表明,在四种不同的 mRNA(长和短 5'UTR 与常规和选择性剪接相结合)中,只有具有短 5'UTR 的常规剪接 mRNA 才能产生全长蛋白。原位杂交和免疫组织化学研究表明,尽管 Brd2 mRNA 在海马体和小脑都有表达,但 Brd2 蛋白只能在小脑浦肯野细胞中检测到,而不能在海马细胞中检测到。这些多层次的调节可能会影响神经发育过程中功能性 BRD2 蛋白的产生,从而影响其在癫痫易感性病因中的作用。