Zhao Zhixing, Lindsay Megan E, Roy Chowdhury Arnab, Robinson Derrick R, Englund Paul T
Department of Biological Chemistry, Johns Hopkins Medical School, Baltimore, MD 21205, USA.
EMBO J. 2008 Jan 9;27(1):143-54. doi: 10.1038/sj.emboj.7601956. Epub 2007 Dec 6.
Kinetoplast DNA (kDNA), the trypanosome mitochondrial genome, is a giant network containing several thousand interlocked DNA rings. Within the mitochondrion, kDNA is condensed into a disk-shaped structure positioned near the flagellar basal body. The disk is linked to the basal body by a remarkable transmembrane filament system named the tripartite attachment complex (TAC). Following kDNA replication, the TAC mediates network segregation, pulling the progeny networks into the daughter cells by their linkage to the basal bodies. So far TAC has been characterized only morphologically with no known protein components. By screening an RNAi library, we discovered p166, a protein localizing between the kDNA and basal body in intact cells and in isolated flagellum-kDNA complexes. RNAi of p166 has only small effects on kDNA replication, but it causes profound defects in network segregation. For example, kDNA replication without segregation causes the networks to grow to enormous size. Thus, p166 is the first reported molecular component of the TAC, and its discovery will facilitate study of kDNA segregation machinery at the molecular level.
动基体DNA(kDNA)是锥虫的线粒体基因组,是一个包含数千个相互连锁的DNA环的巨大网络。在线粒体内,kDNA浓缩成一个位于鞭毛基体附近的盘状结构。该盘通过一个名为三联体附着复合体(TAC)的显著跨膜细丝系统与基体相连。kDNA复制后,TAC介导网络分离,通过与基体的连接将子代网络拉进子细胞。到目前为止,TAC仅在形态学上有特征描述,尚无已知的蛋白质成分。通过筛选RNA干扰文库,我们发现了p166,一种在完整细胞和分离的鞭毛-kDNA复合体中定位于kDNA和基体之间的蛋白质。p166的RNA干扰对kDNA复制只有很小的影响,但它会导致网络分离出现严重缺陷。例如,kDNA复制但不分离会使网络生长到巨大尺寸。因此,p166是首次报道的TAC分子成分,其发现将有助于在分子水平上研究kDNA分离机制。