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Effects of ecdysone and juvenile hormone on the syntheis of specific proteins

蜕皮激素和保幼激素对特定蛋白质合成的影响

基本信息

批准号：
62540541
负责人：
OHTAKI Tetsuya
金额：
$ 1.09万
依托单位：
Kanazawa University, Faculty of Science
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1988
项目状态：
已结题

项目摘要

Effects of ecdysone and jevenile hormone on the synthesis of lectin-like protein, ecdysone receptor protein and prothoracicotropic hormone were studied in larvae and pupae of Bombyx silkworm. The lectin-like protein appeared in hemolymph just before 4th ecdysis and soon after the gut purge in the 5th instar. Change of titer of this protein is almost parallel to the change of ecdysone titer. This protein is of a strong hemagglutinin activity. Using this reaction, the protein was isolated and identified. Molecular weight is about 350KD, and it is thought as a tetramer of 88KD and 90KD polypeptide. Sugers, particularly, glucronic acid, inhibit hemagglutinin activity.Ecdysone receptor protein was isolated from the imaginal wing discs of Bombyx silkworm. Using radioactive Ponasterone A, binding coefficeient to ecdysteroid was determined as 4.1 or 4.3 nM. Molecular weight of the receptor protein was 2.7KD and 6.0KD. To fishing out of the receptor gene from genomic library of the silkworm, a common DNA binding domain of steroid hormones were selected and 15 sequences of common DNA sequence was synthesized. Change of amount of this protein in imaginal discs was studied through 5th instar larve. However, in terms of binding activity/g total protein was almost constant with some fluctiation.Change of ecdysone titer and development of imaginal discs were studied diapuse- and nondiapause-destined culture of the flesh fly. In diapause-destined larvae and prepupae, ecdysone content increased just after pupariation and then decreased to almost undetectable level before pupation. However, in nondiapause-destined larvae second increase of ecdysone content was observed several hours before pupation. Pupal development of imaginal discs was earlier in nondeapause-destined animals than deapause-destined ones, although pupation time was almost same between them.

本文研究了蜕皮激素和促性腺激素对家蚕幼虫和蛹凝集素样蛋白、蜕皮激素受体蛋白和促前胸腺激素合成的影响。在第4龄蜕皮前和第5龄肠道净化后不久，血淋巴中出现了凝集素样蛋白。该蛋白滴度的变化与蜕皮激素滴度的变化几乎平行。该蛋白具有很强的血凝活性。利用该反应，分离并鉴定了蛋白质。分子量约为350 KD，被认为是88 KD和90 KD多肽的四聚体。糖，特别是葡萄糖酸，抑制血凝素活性。蜕皮激素受体蛋白是从家蚕成虫翅盘中分离得到的。使用放射性Ponasterone A，测定与蜕皮甾类的结合系数为4.1或4.3 nM。受体蛋白分子量为2.7KD和6.0KD。为了从家蚕基因组文库中钓出受体基因，选择了一个甾体激素的共同DNA结合域，合成了15个共同DNA序列。以5龄幼虫为材料，研究了该蛋白在成虫盘中的含量变化。通过对肉蝇滞育和非滞育培养，研究了蜕皮激素滴度的变化和成虫盘的发育。滞育幼虫和预蛹期的蜕皮激素含量在化蛹后上升，化蛹前下降到几乎检测不到的水平。然而，在nondiapause-destined幼虫蜕皮激素含量的第二次增加，观察化蛹前几个小时。蛹盘发育在非滞育动物早于滞育动物，虽然化蛹的时间几乎是相同的。