Genomic organization, chromosomal localization, and promoter of human gene for FK506-binding protein 12.6
Genomic organization, chromosomal localization, and promoter of human gene for FK506-binding protein 12.6
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DOI：
10.1016/j.gene.2005.07.004
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发表时间：
2005-10-24
期刊：
影响因子：
3.5
通讯作者：
Okamoto, H
中科院分区：
文献类型：
--
作者：
Nakazawa, T;Takasawa, S;Okamoto, H
环ADP-核糖（cADPR）诱导胰岛微粒体释放Ca~（2+）用于胰岛素分泌。已经证明cADPR与大鼠胰岛兰尼碱受体上的FK506结合蛋白12.6（FKBP 12.6）结合，并且cADPR与FKBP 12.6的结合使兰尼碱受体从FKBP 12.6释放，导致其释放Ca 2 +[Noguchi，N.，Takasawa，S.，Nata，K.，Tohgo，A.，Kato，I，Ikehata，F.，米仓，H.，冈本，H.，1997.环ADP-核糖与FK506结合蛋白结合，从胰岛微粒体释放Ca2+。J. Biol. Chem. 272，3133 - 3136.]。在这项研究中，我们克隆了人FKBP12.6的结构组织，这是高度同源的人FKBP12，并分析了FKBP12.6和FKBP12的启动子。人FKBP12.6基因长度约16 kb，由4个外显子和3个内含子组成。FYBP12.6基因外显子-内含子连接的位置与FKBP12基因完全匹配，除了FKBP12有一个额外的外显子（外显子V）专门编码3'-UTR。荧光原位杂交结果表明，FKBP 12.6基因定位于2号染色体p21 - 23，与FKBP 12基因的定位（20号染色体p13）不同。报告基因分析表明，FKBP 12.6的-58与-24相似的区域和FKBP 12的-106与-79相似的区域对于启动子活性是重要的。启动子包含FKBP 12.6中Sp家族和FK-BP 12中Ets-1的共有转录因子结合序列。电泳迁移率变动分析表明，核蛋白结合的启动子。FKBP12.6启动子上的DNA/蛋白复合物被Sp1共有探针竞争，该复合物被抗Sp3抗体超移位。另一方面，FKBP 12启动子上的DNA/蛋白复合物被Ets-1共有探针竞争而不被其突变探针竞争，表明Sp3和Ets-1分别在FKBP 12.6和FKBP 12的转录中起重要作用。(c)2005 Elsevier B.V.保留所有权利。
Cyclic ADP-ribose (cADPR) induces the release of Ca2+ from microsomes of pancreatic islets for insulin secretion. It has been demonstrated that cADPR binds to FK506-binding protein 12.6 (FKBP 12.6) on rat islet ryanodine receptor and that the binding of cADPR to FKBP 12.6 frees the ryanodine receptor from FKBP 12.6, causing it to release Ca2+ [Noguchi, N., Takasawa, S., Nata, K., Tohgo, A., Kato, I, Ikehata, F., Yonekura, H., Okamoto, H., 1997. Cyclic ADP-ribose binds to FK506-binding protein to release Ca2+ from islet microsomes. J. Biol. Chem. 272,3133 -3136.]. In this study, we cloned, characterized the structural organization of the human FKBP12.6, which is highly homologous to human FKBP12, and analyzed the promoters for FKBP12.6 and FKBP12. Human FKBP12.6 gene spanned about 16 kb in length and consisted of four exons and three introns. The positions of exon-intronjunction of the FYBP12.6 gene were perfectly matched with those of FKBP12 gene except that FKBP12 has an additional exon, exon V, to code exclusively for 3'-UTR. Fluorescence in situ hybridization revealed that the FKBP12.6 gene was located on chromosome 2 p21-23, which is different from the locus (chromosome 20 p13) of the FKBP12 gene. Reporter gene analyses revealed that the regions of - 58 similar to - 24 of FKBP12.6 and - 106 similar to - 79 of FKBP12 are important for promoter activities. The promoters contain a consensus transcription factor binding sequence for Sp family in FKBP12.6 and Ets-1 in FK-BP12. Electrophoretic mobility shift assays showed that nuclear proteins bind to the promoters. The DNA/protein complex on FKBP12.6 promoter was competed out by Sp1 consensus probe and the complex was supershifted by anti-Sp3 antibodies. On the other hand, the DNA/protein complex on FKBP12 promoter was competed out by Ets-1 consensus probe but not by its mutant probe, indicating that Sp3 and Ets-1 play an essential role in transcription of FKBP12.6 and FKBP12, respectively. (c) 2005 Elsevier B.V. All rights reserved.