兔HKalpha2基因启动子的特征分析
Characterization of the rabbit HKalpha2 gene promoter.
作者信息
Zies Deborah L, Gumz Michelle L, Wingo Charles S, Cain Brian D
机构信息
Department of Biochemistry, University of Florida College of Medicine, 1600 SW Archer Road, Gainesville, FL 32610, USA.
出版信息
Biochim Biophys Acta. 2006 Oct;1759(10):443-50. doi: 10.1016/j.bbaexp.2006.08.007. Epub 2006 Sep 12.
The HKalpha2 gene directs synthesis of the HKalpha2 subunit of the H(+), K(+)-ATPase. In the kidney and colon, the gene is highly expressed and is thought to play a role in potassium (K(+)) conservation. The rabbit has been an important experimental system for physiological studies of ion transport in the kidney, so the rabbit HKalpha2 gene has been cloned and characterized. The genomic clones and the previously reported HKalpha2a and HKalpha2c subunit cDNAs provided a means to address several issues regarding the structure and expression of the HKalpha2 gene. First, the genomic organization established that the rabbit HKalpha2 gene was unambiguously homologous to the mouse HKalpha2 gene and the human ATP1AL1 gene. Second, the mapping of the transcription start site for the alternate transcript, HKalpha2c, confirmed that it was an authentic rabbit transcript. Finally, isolation of DNA from the 5' end of the HKalpha2 gene enabled us to initiate studies on its regulation in the rabbit cortical collecting duct. The promoter and two putative negative regulatory regions were identified and the effect of cell confluency on gene expression was studied.
HKalpha2基因指导H(+)、K(+)-ATP酶的HKalpha2亚基的合成。在肾脏和结肠中，该基因高度表达，被认为在钾（K(+)）的保存中起作用。兔子一直是肾脏离子转运生理研究的重要实验系统，因此兔子的HKalpha2基因已被克隆并进行了表征。基因组克隆以及先前报道的HKalpha2a和HKalpha2c亚基cDNA为解决有关HKalpha2基因结构和表达的几个问题提供了一种手段。首先，基因组组织确定兔子的HKalpha2基因与小鼠的HKalpha2基因和人类的ATP1AL1基因明确同源。其次，对交替转录本HKalpha2c的转录起始位点进行定位，证实它是一个真正的兔子转录本。最后，从HKalpha2基因的5'端分离DNA使我们能够启动对其在兔子皮质集合管中的调控的研究。确定了启动子和两个假定的负调控区域，并研究了细胞汇合对基因表达的影响。
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