Exon organization and novel alternative splicing of the human ANK2 gene: Implications for cardiac function and human cardiac disease
Exon organization and novel alternative splicing of the human ANK2 gene: Implications for cardiac function and human cardiac disease
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DOI：
10.1016/j.yjmcc.2008.08.005
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发表时间：
2008-12-01
影响因子：
5
通讯作者：
Mohler, Peter J.
中科院分区：
文献类型：
--
作者：
Cunha, Shane R.;Le Scouarnec, Solena;Mohler, Peter J.
最近的研究结果表明，在正常的心血管生理中，ankle-B功能发挥着关键作用。具体而言，小鼠中的锚蛋白-B表达降低或人类锚蛋白-B基因（ANK 2）突变导致潜在致命的心律失常。尽管ANK 2-B在心脏中的作用是明确的，但ANK 2的转录调控机制尚不清楚。事实上，到目前为止还没有ANK 2基因组结构的描述。本研究的目的是提供一个全面的描述ANK 2基因，并评估在心脏中与ANK 2转录相关的可变剪接事件的相对表达。使用逆转录酶PCR从人类心脏中分离的mRNA，我们确定了7个新的外显子与ANK 2基因，包括一个替代的第一外显子位于类似于145 kb的上游先前确定的第一外显子。此外，我们确定了30多个与ANK 2 mRNA转录相关的选择性剪接事件。使用实时PCR和外显子边界跨越引物来选择性地扩增这些剪接变体，我们证明这些变体在人类心脏中以不同的水平表达。最后，锚蛋白-B免疫印迹分析证实了心脏中锚蛋白-B多肽的异质群体的表达。ANK 2由53个外显子组成，其跨度与人类4号染色体上的560 kb相似。此外，我们的数据表明，ANK 2受到复杂的转录调控，可能导致不同的锚蛋白-B多肽功能。(C)2008年爱思唯尔公司All Rights reserved.
Recent findings illustrate a critical role for ankyrin-B function in normal cardiovascular physiology. Specifically, decreased expression of ankyrin-B in mice or human mutations in the ankyrin-B gene (ANK2) results in potentially fatal cardiac arrhythmias. Despite the clear role of ankyrin-B in heart, the mechanisms underlying transcriptional regulation of ANK2 are unknown. In fact, to date there is no description of ANK2 genomic organization. The aims of this study were to provide a comprehensive description of the ANK2 gene and to evaluate the relative expression of alternative splicing events associated with ANK2 transcription in heart. Using reverse-transcriptase PCR on mRNA isolated from human hearts, we identify seven new exons associated with the ANK2 gene including an alternative first exon located similar to 145 kb upstream of the previously-identified first exon. In addition, we identify over thirty alternative splicing events associated with ANK2 mRNA transcripts. Using real-time PCR and exon boundary-spanning primers to selectively amplify these splice variants, we demonstrate that these Variants are expressed at varying levels in human heart. Finally, ankyrin-B immunoblot analysis demonstrates the expression of a heterogeneous population of ankyrin-B polypeptides in heart. ANK2 consists of 53 exons that span similar to 560 kb on human chromosome 4. Additionally, our data demonstrates that ANK2 is subject to complex transcriptional regulation that likely results in differential ankyrin-B polypeptide function. (C) 2008 Elsevier Inc. All Rights reserved.