Studies on structure and function of organelles dividing machines based on genome information

为了阐明线粒体和质体在细胞核控制下的分裂机制，我们以原始红藻Cyanidioschyzon merolae为材料。墨藻细胞具有非常有利的细胞器分裂，因为细胞中含有最少数量的细胞器，这些细胞器在光照和黑暗周期中同步分裂。所得数据与高等生物的数据进行了比较。由于PD环的周长是线粒体分裂环的2.5倍，我们首先检测了质体分裂环。为了从完整的PD环装置中获得纯蛋白质，我们改进了叶绿体分裂同步效率、分离纯分裂叶绿体、通过各种洗涤处理分离完整的PD环装置以及从分离的完整PD环装置中完全提取蛋白质等几个方面。利用二维电泳获得SDS凝胶点，并利用MALDI-TOFMS对斑点进行分析。我们鉴定了30个与PD环部分相关的基因，并制备了它们的抗体。我们在PD环装置中发现了至少4个新的基因，并根据现有的方法继续发现更多与细胞器分裂密切相关的基因。

To elucidate the mechanism of mitochondria and plastid division under the control of the cell nucleus, we provided the primitive red alga Cyanidioschyzon merolae as a material. C.merolae cells have very advantageous for organelle divisions because the cell contains minimum number of organelles of which divisions were synchronized by light and dark cycles. The obtained data are compared with those of higher organisms. We first examined plastid dividing (PD) ring apparatus as the circumference of PD ring was 2.5 times longer than that of mitochondrial dividing (MD) ring apparatus. To obtain pure proteins from intact PD ring apparatus, we improved several points such as efficiency of synchronization of chloroplast division, isolation of pure dividing chloroplasts, isolation of intact PD ring apparatus by various detergent treatments and complete extraction of proteins from isolated intact PD ring apparatus. We then obtained SDS gel spots by using two dimensional electrophoresis and analyzed the spots by MALDI-TOFMS on basis of C.merolae genome information. We identified about 30 genes related to PD ring fractions and made their antibodies. We found at least novel 4 genes which are contained in PD ring apparatus and continue to find more genes related intimately to organelle divisions according to the established methods.