Molecular characterization and functional expression of the DSC1 channel.

Molecular characterization and functional expression of the DSC1 channel.
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DOI:
10.1016/j.ibmb.2011.04.010
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发表时间:
2011-07
影响因子:
3.8
通讯作者:
Dong, Ke
Dong, Ke
中科院分区:
农林科学2区
文献类型:
--
作者:
Zhang, Tianxiang;Liu, Zhiqi;Song, Weizhong;Du, Yuzhe;Dong, Ke

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果蝇钠离子通道1(DSC 1)与脊椎动物和无脊椎动物钠离子通道基因具有高度的序列相似性,被预测为编码一种钠离子通道。然而,BSC 1,DSC 1直系同源物在德国小蠊,最近被证明编码一个阳离子通道与离子选择性对Ca 2+。在本研究中,我们分离了20个全长cDNA克隆,涵盖了整个编码区的DSC 1基因从D。通过逆转录-聚合酶链反应(RT-PCR)检测黑腹果蝇的DNA。20个克隆的序列分析揭示了9个可选的外显子,其中4个包含框内终止密码子;和13个潜在的A到I RNA编辑位点。由于独特的RNA编辑,这20个克隆可以分为8种剪接类型,代表20种不同的转录本。三个变体产生的DSC 1电流在非洲爪蟾卵母细胞中表达。与BSC 1通道一样,所有三个功能性DSC 1通道都对Ca 2+和Ba 2+具有渗透性,在没有外部Ca 2+的情况下也对Na+具有渗透性。此外,DSC 1通道对河豚毒素(一种有效且特异性的钠通道阻断剂)不敏感。我们的研究表明,DSC 1编码的电压门控阳离子通道类似于B中的BSC 1通道。德国的。广泛的选择性剪接和RNA编辑的DSC 1的成绩单表明DSC 1通道的分子和功能的多样性。
Drosophila Sodium Channel 1 (DSC1) was predicted to encode a sodium channel based on a high sequence similarity with vertebrate and invertebrate sodium channel genes. However, BSC1, a DSC1 ortholog in Blattella germanica, was recently shown to encode a cation channel with ion selectivity towards Ca2+. In this study, we isolated a total of 20 full-length cDNA clones that cover the entire coding region of the DSC1 gene from adults of D. melanogaster by reverse transcription-polymerase chain reaction. Sequence analysis of the 20 clones revealed nine optional exons, four of which contain in-frame stop codons; and 13 potential A-to-I RNA editing sites. The 20 clones can be grouped into eight splice types and represent 20 different transcripts because of unique RNA editing. Three variants generated DSC1 currents when expressed in Xenopus oocytes. Like the BSC1 channel, all three functional DSC1 channels are permeable to Ca2+ and Ba2+, and also to Na+ in the absence of external Ca2+. Furthermore, the DSC1 channel is insensitive to tetrodotoxin, a potent and specific sodium channel blocker. Our study shows that DSC1 encodes a voltage-gated cation channel similar to the BSC1 channel in B. germanica. Extensive alternative splicing and RNA editing of the DSC1 transcripts suggest the molecular and functional diversity of the DSC1 channel.
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