The mouse oviduct-specific glycoprotein gene : Genomic organization and structure of the 5'-flanking regulatory region

小鼠输卵管特异性糖蛋白基因：5-侧翼调节区的基因组组织和结构

基本信息

批准号：
09671663
负责人：
ARAKI Yoshihiko
金额：
$ 1.86万
依托单位：
Yamagata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671663/
关键词：
mouse oviduct glycoprotein genomic cloning estrogen responsive element プロモーター

项目摘要

A member of the chitinase protein family, oviduct-specific glycoprotein (OGP), can directly associate with gametes or the early embryo in the oviduct. Although the glycoprotein is widely distributed among mammalian species and there is indirect evidence concerning the involvement of the molecule for fertilization process, its physiological functions are far from completely understood. To understand the fundamental mechanisms that direct gene expression as well as to know the physiological significance of OGP, we have isolated and characterized a mouse OGP gene(mogp-1). The gene was found to span 13.4-kilo bases (kb) including 11 exons and 10 introns. The genomic organization of mogp-1 is well conserved compared to the other members of the chitinase family. Two transcription initiation sites were found at positions 18 and 14 upstream from the first ATG codon. Fluorescence in situ hybridization analysis demonstrated that the mogp-1 was located the on R-positive F3 band of mouse chromosome 3. Although the putative promoter region of mogp-1 lacked typical TATA, CAAT or GC box sequences, the region contained several motif sequences of transcription factor binding sites including 10 half-palindromic estrogen responsive elements (ERE) and an imperfect ERE. Transient transfection experiments demonstrated that promoter activity could be modulated by various sequences within the 2.2-kb of the 5'-flanking region, and that the mogp-1 promoter was transactivated in an estrogen receptor positive cell line, MCF-7, by the addition of 17beta-estradiol (E2). In addition, a relevant promoter activity for E2 responsiveness resides within the first 270 bp upstream of the mogp-1. These findings should facilitate our understanding of the regulation of OGP gene expression and they may be helpful for designing experiments to unravel the role of OGP in the process of mammalian fertilization.

几丁质蛋白蛋白家族的成员Oviduct特异性糖蛋白（OGP）可以直接与配子或产卵中的早期胚胎相关联。尽管糖蛋白广泛分布在哺乳动物物种之间，并且有关于该分子施肥过程中参与的间接证据，但其生理功能远非完全了解。为了了解直接基因表达以及了解OGP的生理意义的基本机制，我们已经分离并表征了小鼠OGP基因（MOGP-1）。发现该基因跨越了13.4 kilo碱基（Kb），包括11个外显子和10个内含子。与几丁质酶家族的其他成员相比，MOGP-1的基因组组织是保守的。在第一个ATG密码子的上游位置发现了两个转录启动位点。荧光原位杂交分析表明，MOGP-1位于小鼠染色体3的R阳性F3带上。尽管MOGP-1的推定启动子区域缺乏典型的TATA，CAAT或GC盒序列，但该区域包含几个Motif序列转录因子结合位点的结合位点，包括10个半圆锥体雌激素反应元件（ERE）和不完美的ERE。瞬态转染实验表明，启动子活性可以通过5'FANKING区域的2.2-kb内的各种序列调节，并且MOGP-1启动子在雌激素受体正阳性细胞系MCF-7中通过添加剂进行反式激活。 17beta-雌二醇（E2）。此外，在MOGP-1的前270 bp中，E2响应能力的相关启动子活性也存在。这些发现应促进我们对OGP基因表达的调节的理解，它们可能有助于设计实验以揭示OGP在哺乳动物受精过程中的作用。