Backert S, Dörfel P, Lurz R, Börner T
Institut für Biologie, Humboldt-Universität zu Berlin, Germany.
Mol Cell Biol. 1996 Nov;16(11):6285-94. doi: 10.1128/MCB.16.11.6285.
The mitochondrial genomes of higher plants are larger and more complex than those of all other groups of organisms. We have studied the in vivo replication of chromosomal and plasmid mitochondrial DNAs prepared from a suspension culture and whole plants of the dicotyledonous higher plant Chenopodium album (L.). Electron microscopic studies revealed sigma-shaped, linear, and open circular molecules (subgenomic circles) of variable size as well as a minicircular plasmid of 1.3 kb (mp1). The distribution of single-stranded mitochondrial DNA in the sigma structures and the detection of entirely single-stranded molecules indicate a rolling-circle type of replication of plasmid mp1 and subgenomic circles. About half of the sigma-like molecules had tails exceeding the lengths of the corresponding circle, suggesting the formation of concatemers. Two replication origins (nicking sites) could be identified on mpl by electron microscopy and by a new approach based on the mapping of restriction fragments representing the identical 5' ends of the tails of sigma-like molecules. These data provide, for the first time, evidence for a rolling-circle mode of replication in the mitochondria of higher plants.
高等植物的线粒体基因组比所有其他生物类群的线粒体基因组更大且更复杂。我们研究了从双子叶高等植物藜(Chenopodium album (L.))的悬浮培养物和整株植物中制备的染色体线粒体DNA和质粒线粒体DNA的体内复制情况。电子显微镜研究揭示了大小可变的σ形、线性和开放环状分子(亚基因组环)以及一个1.3 kb的小环状质粒(mp1)。单链线粒体DNA在σ结构中的分布以及完全单链分子的检测表明质粒mp1和亚基因组环的复制方式为滚环复制。大约一半的σ样分子有超过相应环长度的尾巴,这表明形成了多联体。通过电子显微镜以及一种基于对代表σ样分子尾巴相同5'末端的限制性片段进行图谱分析的新方法,可以在mpl上鉴定出两个复制起点(切口位点)。这些数据首次为高等植物线粒体中的滚环复制模式提供了证据。