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.

本研究项目旨在获得线索，以了解海胆早期发育过程中基因在时间和空间上表达的机制。为此，我们选择了一个在海胆胚胎中以时间和空间调节方式表达的芳基硫酸酯酶(Ars)基因。海胆Ars基因在其基本启动子上有一个TATA盒，在其5‘侧翼区域有几个G-弦序列和一个长的多嘧啶区域。本研究取得了以下结果。(2)以人工合成的聚(DG)-聚(DC)为探针，通过西南杂交获得了4个G-链DNA结合蛋白的cDNA克隆。所有这些基因在其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之间的一小段决定。-100和-252位之间的两个顺式元件似乎与ARS基因表达的时间调控有关。(4)第一内含子中的229nts序列显著增强了这些元素的活性。这些结果都是通过荧光素酶分析获得的。(5)构建了体外高效转录ARS基因的核提取系统。

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