Requlation of gene expression in sea urchin embryo

海胆胚胎基因表达调控

• 批准号: 05454653
• 负责人: SHIMADA Hiraku
• 金额: $ 4.22万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05454653/
• 关键词: sea urchin; development; gene expression; arylsufatase; cis-element; DNA binding protein; G-string; luciferase assay; ウニ発生; アリールスルファターゼ; 転写調節; 遺伝子発現; 転写因子; インビトロ転写

This research project has been proposed to obtain clues to understand a mechanism underlying temporal and spatial expression of genes during early development of sea urchin. For this purpose we selected a gene for arylsulfatase (Ars) that expresses in the temporally and spatially regulated manner in sea urchin embryo. The sea urchin Ars gene contains a TATA box in its basal promoter, several G-string sequences and a long polypyrimidine region in its 5' flanking region. Following results were obtained in this study. (1) The long polypyrimidine region can form a triplex DNA configurationthrough Hoogsten base parings, (2) four cDNA clones for G-string DNA-binding proteins were obtained by south-western blotting using a synthetic poly (dG)-poly(dC) as the probe. All these cDNAs contain GA-rich sequence in its ORF.(3) Temporal expression of the Ars gene is primarily determined by a short region between -1 and -100b positions. Two cis-elements between -100 and -252 position seem to be related to temporal regulation of the Ars gene expression. (4) A 229nts sequence in the first intron strongly enhances the activity of these elements. all these results were obtained by luciferase assays. (5) A nuclear extract system to effectively transcribe the Ars gene in vitro was constructed.

该研究项目旨在获得线索，以了解海胆早期发育过程中基因时空表达的机制。为此，我们选择了一个芳基硫酸酯酶（Ars）的基因，在海胆胚胎中的时间和空间调控的方式表达。海胆Ars基因在其基础启动子中包含一个TATA盒，在其5'侧翼区包含几个G串序列和一个长多嘧啶区。本研究获得了以下结果。(1)（2）以人工合成的poly（dG）-poly（dC）为探针，经South-western印迹杂交，获得了4个G-string DNA结合蛋白的cDNA克隆。所有这些cDNA的ORF中都含有富含GA的序列。(3)Ars基因的时间表达主要由-1和-100b位置之间的短区域决定。在-100和-252位之间的两个顺式元件似乎与Ars基因表达的时间调节有关。(4)第一个内含子中的229 nts序列强烈增强了这些元件的活性。所有这些结果都是通过荧光素酶测定获得的。(5)构建了Ars基因体外转录的核提取体系。

项目成果

T.Yamamoto et al.: "A long polypyrimidine:polypurine seguence in 5'flanking region of arylsulfatase gene of sea urchin embryo." International Journal of Developmental Biology. 38. 337-344 (1994)

T.Yamamoto 等人：“海胆胚胎芳基硫酸酯酶基因 5 侧翼区域的长聚嘧啶：聚嘌呤序列。”

Y.Iuchi et al.: "Detection and characterization of the cis-element in the first intron of the Ars gene in sea urchin" Development,Growth & Differentiation. 37(in press). (1995)

Y.Iuchi 等人：“海胆 Ars 基因第一个内含子中顺式元件的检测和表征”发育、生长

K.Akasaka et al: "Corrected structure of the 5′flanking region of arylsulfatase gene of the sea urchin,Hemicentrotus pulcherrimus." Development,Growth & Differentiation. 36. 633-636 (1994)

K.Akasaka 等人：“海胆 Hemicentrotus pulcherrimus 芳基硫酸酯酶基因 5 侧翼区域的校正结构。” 发育、生长和分化 36. 633-636 (1994)。

K.Akasaka, N.Sakamoto, T.Yamamoto, J.Morokuma, F.Fujikawa, K.Takata, S.Eguchi and H.Shimada: "Corrected structure of the 5' flanking region of arylsulfatase gene of the sea urchin, Hemicentrotus pulcherrimus" Develop.Growth & Differ.36. 633-636 (1994)

K.Akasaka、N.Sakamoto、T.Yamamoto、J.Morokuma、F.Fujikawa、K.Takata、S.Eguchi 和 H.Shimada：“海胆 Hemicentrotus 芳基硫酸酯酶基因 5 侧翼区域的校正结构

Y.Iuchi, J.Morokuma, K,Akasaka and H.Shimada: "Detection and characterization of the cis-element in the first intron of the Ars gene in sea urchin" Develop.Growth & Differ.37 (in press). (1995)

Y.Iuchi、J.Morokuma、K、Akasaka 和 H.Shimada：“海胆 Ars 基因第一个内含子中顺式元件的检测和表征” Develop.Growth