STUDY ON TRANSCRIPTION FACTORS REGULATING GENE EXPRESSION IN SEA URCHIN EMBRYOS

海胆胚胎基因表达调控转录因子的研究

• 批准号: 07458195
• 负责人: SHIMADA Hiraku
• 金额: $ 5.12万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07458195/
• 关键词: sea urchin; arylsulfatase; gene expression; embryo; enhancer; 0tx; insulator; 発生; 転写因子; ミトコンドリア; リボソームRNA; Ars遺伝子; 転写調節; Gストリング

The present research project has been undertaken to elucidate a cascade of transcription factors related to temporally and spatially regulated expression of genes during early development of sea urchin. Results obtained during the research period for this grant are as follows.(1) A procedure to introduce exogenous DNA into sea urchin eggs by a particle gun has been developed.(2) Temporal expression of the Ars gene is regulated by two distinct cisactive elements in its promoter region (from -252 to +38bp), though level of transcription initiated by these elements alone is very low and declines rapidly.(3) GGATTA sequences (known as binding sites for transcription factor 0tx) in the first intron of the Ars gene act as a transcriptional enhancer of this gene. Two distinct cDNAs (Hp0tx_E and Hp0tx_L) encoding Hp0tx have been cloned. Hp0tx_E is a maternal factor and suppresses the Ars gene transcription, while Hp0tx_L is produced in hatched blastulae and activates the Ars gene transcription. Two cDNAs are produced from the same gene by alternative splicing.(4) It is found that the region from -2686 to -2113bp of the Ars gene acts as an transcriptional insulator that prevents the Ars gene to be transcribed under the influence of promoter of the gene located in a closely vicinity.

本研究旨在阐明海胆早期发育过程中与基因时空调控表达相关的一系列转录因子。在该补助金的研究期间取得的成果如下。(1)利用粒子枪将外源DNA导入海胆卵。(2)Ars基因的时间表达受其启动子区域（从-252到+38bp）的两个不同的顺式作用元件调节，尽管单独由这些元件启动的转录水平非常低并且迅速下降。(3)在Ars基因的第一个内含子中的GGATTA序列（称为转录因子0 tx的结合位点）充当该基因的转录增强子。已克隆了编码Hp 0 tx的两个不同cDNA（Hp0tx_E和Hp0tx_L）。Hp0tx_E是一种母源性因子，抑制Ars基因的转录，而Hp0tx_L在孵化囊胚中产生，激活Ars基因的转录。通过选择性剪接从同一基因产生两个cDNA。(4)发现Ars基因的-2686 ~-2113bp区域是一个转录绝缘子，在邻近基因启动子的影响下，阻止Ars基因的转录。

项目成果

J.Morokuma, K.Akasaka, K.Mitsunaga-Nakatsubo and H.Shimada: "A cis-regulatory element within the 5' flanking region of arylsulfatase gene of sea urchin, Hemicentrotus pulcherrimus." Develop.Growth & Differ. 39(in press). (1997)

J.Morokuma、K.Akasaka、K.Mitsunaga-Nakatsubo 和 H.Shimada：“海胆 Hemicentrotus pulcherrimus 芳基硫酸酯酶基因 5 侧翼区域内的顺式调控元件。”

N. Sakamoto et al: "Atriplex DNA configuration of the polypyrimidine: polypurine stretch in the 5′ flanking region of the sea urchin arylsulfatase gene." Zoological Science. 13. 105-109 (1996)

N. Sakamoto 等人：“聚嘧啶的 Atriplex DNA 构型：海胆芳基硫酸酯酶基因 5 侧翼区域的聚嘌呤延伸。动物学科学”13. 105-109 (1996)。

Harada et al: "Spatial expression of a forkhead homologue in the sea urchin embryo." Mechanism of Development. 60. 163-173 (1996)

Harada 等人：“叉头同源物在海胆胚胎中的空间表达。”

N.Sakamoto, K.Akasaka, T.Yamamoto and H.Shimada: "A triplex DNA configuration of the polypyrimidine : polypurine stretch in the 5' flanking region of the sea urchin arylsulfatase gene." Zool.Sci.13. 105-109 (1996)

N.Sakamoto、K.Akasaka、T.Yamamoto 和 H.Shimada：“聚嘧啶的三链 DNA 构型：海胆芳基硫酸酯酶基因 5 侧翼区域的聚嘌呤延伸。”

Sakamoto et al.: "A triplex DNA configuration of the polypyrimldine : polypurine stretch in the 5' flanking region of the sea urchin arylsulfatase gene." Zoological Science. 13. 105-109 (1996)

Sakamoto 等人：“聚嘧啶的三链 DNA 构型：海胆芳基硫酸酯酶基因 5 侧翼区域的聚嘌呤延伸。”