Sak kinase gene structure and transcriptional regulation
Sak kinase gene structure and transcriptional regulation
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DOI：
10.1016/s0378-1119(99)00467-9
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发表时间：
2000-01-04
期刊：
影响因子：
3.5
通讯作者：
Dennis, JW
中科院分区：
文献类型：
--
作者：
Hudson, JW;Chen, LY;Dennis, JW
Sak基因编码丝氨酸/苏氨酸激酶，其是有丝分裂调节因子的波罗家族的成员。Sak转录本存在于S/G2/M期细胞以及小鼠胚胎和成体组织的增殖细胞层中。在这份报告中，我们已经确定了小鼠Sak基因的结构，Sak染色体的位置，并确定了启动子。通过RFLP分析确定，鼠Sak基因位于小鼠13号染色体的近端臂上。鼠基因包含跨越16 kb基因组序列的15个编码外显子，并且编码两个交替剪接的转录物。Sak-a是主要的转录本，由15个外显子编码，而在外显子5和6处的转录和选择性剪接的早期终止导致Sak-b。该截短的转录物编码完整的激酶结构域和从与外显子5邻接的147 bp序列翻译的羧基末端。据报道，人Sak-a（Stk 18）cDNA含有对应于小鼠Sak-b尾的插入序列。小鼠Sak的引物延伸分析揭示了相对于翻译起始的位置-303bp处的一个主要转录起始位点。位于Sak转录起始点5'的3.5 kb基因组片段以方向依赖性方式驱动CHO和GC 1-SPG细胞中的内切酶报告基因的表达。使用各种Sak启动子/荧光素酶构建体，表达所需的核心启动子区域位于信使Cap位点的400 bp内，并且序列进一步5'强烈抑制转录。(C)2000 Elsevier Science B. V.保留所有权利。
The Sak gene encodes a serine/threonine kinase, which is a member of the Polo family of mitotic regulators. Sak transcripts are present in S/G2/M phase cells, and in proliferating cell layers of the mouse embryo and adult tissues. In this report, we have characterized the murine Sak gene structure, the Sak chromosomal location, and identified the promoter. The murine Sak gene is located on the proximal arm of mouse chromosome 13, as determined by RFLP analysis. The murine gene comprises 15 coding exons spanning 16 kb of genomic sequence, and encodes two alternately spliced transcripts. Sak-a, the predominant transcript, is encoded by 15 exons, while early termination of transcription and alternative splicing at exons 5 and 6 results in Sak-b. This truncated transcript encodes the complete kinase domain and a carboxyl end translated from 147 bp of sequence contiguous with exon 5. Human Sak-a (Stk18) cDNA is reported to contain an insertion of sequence corresponding to the mouse Sak-b tail. Primer extension analysis of murine Sak revealed one major transcription start site at position -303 bp relative to the start of translation. A genomic fragment of 3.5 kb located 5' of the Sak transcriptional start drives expression of a luciferase-reporter gene in CHO and GC1-SPG cells in an orientation-dependent fashion. Using various Sak promoter/luciferase constructs, the core promoter region required for expression was located within 400 bp of the message Cap site, and sequence further 5' strongly suppressed transcription. (C) 2000 Elsevier Science B.V. All rights reserved.