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Study on the mechanism in the cell cycle synchronization in the algal multinucleate cells

藻类多核细胞细胞周期同步机制的研究

基本信息

批准号：
05640743
负责人：
MOTOMURA Taizo
金额：
$ 1.41万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
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

项目摘要

Multinucleate cells have been reported in many organisms, including fungi, animals and plants.It is well known that the cell cycle (or, proceeding of DNA synthesis and mitosis) of each nucleus in the multinucleate cells pregresses synchronously.While, in multinucleate green algae, several types of mitotic pattern can be observed.In Bryopsis, each nucleus divides independently.Almost synchronous nuclear division occurs in discrete patches in Boergesenia.And in Acrosiphonia, almost all nuclei in the cell divide synchronously.When considering the cell cycle regulation by p34^<cdc2> and cyclins, these phenomena in the multinucleate green algae are unusual.In this study.I tried to examine the mechanism in the cell cycle regulation in the green algae using cytological and molecular biological techniques.Especially, a cDNA clone of Acrosiphonia that is homologous to the cdc2 genes was isolated using PCR,and its nucleotide and deduced amino acid sequences was determined.Putative amino acid sequence of Acrosiphonia cdc2 gene showed 47.2% homology to yeast cdc2 protein kinase.When analyzed with a monoclonal antibody (PSTAIRE antibody) against a cdc2 protein kinase, a band of Mr38,000 was recognized.Protein kinase encoded in cdc2 gene of the eukaryotes has always ahown Mr34,000, therefore, the molecular weight of Acrosiphonia cdc2 protein kinase was very unique.Adding to it, the antibody could not detect any protein in Bryopsis and Boergesenia.These results might suggest that the regulation in the cell cycle of the multinucleate green algae was carried out by considerably deviated cdc2 gene product from other eukaryotes.

多核细胞在真菌、动物和植物等生物中都有报道。众所周知，多核细胞中每一个核的细胞周期(或DNA合成和有丝分裂)是同步进行的。而在多核绿藻中，可以观察到几种类型的有丝分裂模式。在苔藓中，每个核独立分裂。在羊栖菜中，几乎同步的核分裂发生在离散的斑块中。在海藻中，几乎所有的核都是同步分裂的。当考虑到p34和lt；cdc2和GT；本研究试图用细胞学和分子生物学技术来研究绿藻细胞周期调控的机制。特别是通过聚合酶链式反应分离到一个与cdc2基因同源的顶管藻属的cDNA克隆，并测定了它的核苷酸和推导的氨基酸序列。顶管藻属cdc2基因的氨基酸序列与酵母cdc2蛋白激酶的氨基酸序列有47.2%的同源性。用抗cdc2蛋白激酶的单抗(PSTAIRE抗体)分析发现，在真核细胞的cdc2基因编码的蛋白激酶中有一条Mr 38000的条带。因此，该基因的氨基酸序列与酵母cdc2蛋白激酶的同源性为47.2%。用抗cdc2蛋白激酶的单抗(PSTAIRE抗体)分析发现，在真核细胞的cdc2基因中编码的蛋白激酶始终有34000个左右Acrosiphia cdc2蛋白激酶的相对分子质量非常独特，在苔藓和白僵菌中没有检测到任何蛋白质，这一结果可能表明多核绿藻细胞周期的调控是通过cdc2基因产物与其他真核生物有较大差异来进行的。