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

为了阐明细胞核控制下线粒体和质体分裂的机制，我们以原始红藻蓝藻Merolae为材料。冬虫夏草细胞对细胞器的分裂非常有利，因为该细胞含有最少数量的细胞器，这些细胞器的分裂是由光和暗周期同步的。将获得的数据与高等生物的数据进行了比较。我们首先检查了叶绿体分裂环(PD环)，因为PD环的周长是线粒体分裂环(MD)环的2.5倍。为了从完整的Pd环器中获得纯净的蛋白质，我们改进了叶绿体分裂的同步化效率、分离纯分裂的叶绿体、用各种去污剂处理分离完整的Pd环器以及从分离的完整Pd环器中完全提取蛋白质。然后利用双向电泳法获得了SDS凝胶斑点，并根据斑点的基因组信息对斑点进行了MALDI-TOFMS分析。我们鉴定了大约30个与PD环组分相关的基因，并制作了它们的抗体。根据已建立的方法，我们至少发现了4个新的包含在PD环器中的基因，并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.