Complete genomic organization and promoter analysis of the round-spotted pufferfish JAK1, JAK2, JAK3, and TYK2 genes.
Complete genomic organization and promoter analysis of the round-spotted pufferfish JAK1, JAK2, JAK3, and TYK2 genes.
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DOI：
10.1089/10445490050085924
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发表时间：
2000-07
影响因子：
3.1
通讯作者：
J. Leu;Shian Jang Yan;Tzuu-fen Lee;Chih Ming Chou;S. Chen;Pung-Pung Hwang-Pung;Chen-Kung Chou;C. Huang
中科院分区：
文献类型：
--
作者：
J. Leu;Shian Jang Yan;Tzuu-fen Lee;Chih Ming Chou;S. Chen;Pung-Pung Hwang-Pung;Chen-Kung Chou;C. Huang
我们以前报道过从圆斑河豚中分离出JAK1基因。在本研究中，我们克隆并鉴定了河豚JAK2、JAK3和TYK2的基因组序列，它们是JAK家族的其他成员。据我们所知，这是首次证实鱼类中存在四个JAK基因的报道。除JAK1外，所有河豚JAK基因均由24个外显子组成；JAK1有一个额外的外显子。比较这些基因的外显子-内含子结构，发现JAK基因的剪接位点几乎相同。此外，所有河豚JAK基因在5'非翻译区都有一个内含子。综合起来，这些数据表明河豚的JAK基因可能是从一个共同的祖先进化而来的。通过cDNA末端的5'快速扩增和序列分析，我们推断出所有JAK基因的启动子区域，发现它们不包含典型的TATA或CCAAT盒，而是包含许多其他潜在的转录因子结合位点。有趣的是，TYK2基因与CDC37以首尾相连的方式与292bp的小基因间区域相连。在这个区域内，有两个转录因子如c-Myb和NF-IL6的潜在结合位点。以CAT或lacZ作为报告基因，在鲤鱼CF细胞系或斑马鱼胚胎中检测了所有JAK基因的推定启动子区域。两种方法都证实了这些启动子在体内和体外的转录活性。
We have previously reported the isolation of the JAK1 gene from the round-spotted pufferfish. In the present study, we cloned and characterized genomic sequences encoding pufferfish JAK2, JAK3, and TYK2, which are other members of JAK family. To our knowledge, this is the first report to demonstrate the existence of four JAK genes in fish. All pufferfish JAK genes except JAK1 are composed of 24 exons; JAK1 has an additional exon. A comparison of the exon-intron organization of these genes revealed that the splice sites of JAK genes are nearly identical. In addition, all pufferfish JAK genes have one intron in the 5' untranslated region. Taken together, these data suggest that the pufferfish JAK genes may have evolved from a common ancestor. By 5' rapid amplification of cDNA ends and sequence analysis, we deduced the promoter regions for all JAK genes and found they do not contain typical TATA or CCAAT boxes but rather numerous other potential binding sites for transcription factors. Interestingly, the TYK2 gene is linked to CDC37 in a head-to-tail manner with a small intergenic region of 292 bp. Within this region, there are two potential binding sites for transcriptional factors such as c-Myb and NF-IL6. The putative promoter regions of all JAK genes were tested either in a carp CF cell line or in zebrafish embryos using CAT or lacZ as reporter genes. Both assays confirmed the transcriptional activities of these promoters in vitro and in vivo.