Molecular cloning and analysis of the mouse gicerin gene
Molecular cloning and analysis of the mouse gicerin gene
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DOI：
10.1016/j.neuint.2004.12.006
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发表时间：
2005-05-01
影响因子：
4.2
通讯作者：
Taira, E
中科院分区：
文献类型：
--
作者：
Kohama, K;Tsukamoto, Y;Taira, E
Gicerin是一种细胞粘附分子，其在细胞外结构域中具有五个免疫球蛋白样环结构，随后是单个跨膜结构域和短的胞质尾。我们已经报道了gicerin参与神经系统的神经延伸和结构组织，并且其在神经系统中的表达在发育期间是高的，并且在出生后急剧降低。为了阐明gicerin的表达调控机制，我们进行了小鼠gicerin的基因组克隆。该基因组克隆全长16 kbp，其中gicerin基因全长8 kbp，由16个外显子组成，上游6 kbp。基因组克隆显示，两种异构体的gicerin产生的选择性剪接外显子15的结果在细胞质结构域组成的63或21个氨基酸。对于gicerin的表达调控，我们发现在PC 12细胞中gicerin的mRNA含量受cAMP的调控。定量PCR分析表明，毛喉素诱导gicerin mRNA的四倍增加。为了表征其启动子区域的参与，我们通过内切酶-报告基因测定法检测了PC 12细胞中的启动子活性。我们发现，位于gicerin基因上游60 bp的CRE位点是导致毛喉素诱导gicerin mRNA表达增加的主要原因。(c)2005爱思唯尔有限公司保留所有权利。
Gicerin is a cell adhesion molecule, which has five immunoglobulin-like loop structures in an extracellular domain followed by a single transmembrane domain and a short cytoplasmic tail. We have reported that gicerin participates in nuerite extension and structural organization of the nervous system, and its expression in the nervous system is high during the development and dramatically decreased after birth. To elucidate the mechanism how the expression of gicerin is regulated, we performed a genomic cloning of a mouse gicerin. A fragment of 16 kbp genomic clone contained 8 kbp gicerin gene composed of 16 exons with 6 kbp upstream region. Genomic cloning revealed that two isoforms of gicerin were generated by an alternative splicing of exon 15 results in cytoplasmic domains composed of either 63 or 21 amino acids. As for an expressional regulation of gicerin, we found that the mRNA content of gicerin in PC12 cells was regulated by cAMP. Quantitative-PCR analysis revealed that forskolin induced four-fold increase of gicerin mRNA. To characterize the involvement of its promoter region, we examined the promoter activity in PC 12 cells by a luciferase-reporter assay. We found that a CRE site located at 60 bp upstream of gicerin gene was responsible for the increase of its mRNA induced by forskolin. (c) 2005 Elsevier Ltd. All rights reserved.