Zuo Xiao Ming, Clark-Walker G Desmond, Chen Xin Jie
Molecular Genetics and Evolution Group, Research School of Biological Sciences, The Australian National University, Canberra, ACT 2601, Australia.
Genetics. 2002 Apr;160(4):1389-400. doi: 10.1093/genetics/160.4.1389.
The Saccharomyces cerevisiae MGM101 gene encodes a DNA-binding protein targeted to mitochondrial nucleoids. MGM101 is essential for maintenance of a functional rho(+) genome because meiotic segregants, with a disrupted mgm101 allele, cannot undergo more than 10 divisions on glycerol medium. Quantitative analysis of mtDNA copy number in a rho(+) strain carrying a temperature-sensitive allele, mgm101-1, revealed that the amount of mtDNA is halved each cell division upon a shift to the restrictive temperature. These data suggest that mtDNA replication is rapidly blocked in cells lacking MGM101. However, a small proportion of meiotic segregants, disrupted in MGM101, have rho(-) genomes that are stably maintained. Interestingly, all surviving rho(-) mtDNAs contain an ori/rep sequence. Disruption of MGM101 in hypersuppressive (HS) strains does not have a significant effect on the propagation of HS rho(-) mtDNA. However, in petites lacking an ori/rep, disruption of MGM101 leads to either a complete loss or a dramatically decreased stability of mtDNA. This discriminatory effect of MGM101 suggests that replication of rho(+) and ori/rep-devoid rho(-) mtDNAs is carried out by the same process. By contrast, the persistence of ori/rep-containing mtDNA in HS petites lacking MGM101 identifies a distinct replication pathway. The alternative mtDNA replication mechanism provided by ori/rep is independent of mitochondrial RNA polymerase encoded by RPO41 as a HS rho(-) genome is stably maintained in a mgm101, rpo41 double mutant.
酿酒酵母MGM101基因编码一种靶向线粒体类核的DNA结合蛋白。MGM101对于维持功能性的ρ(+)基因组至关重要,因为带有mgm101等位基因破坏的减数分裂分离株在甘油培养基上不能进行超过10次分裂。对携带温度敏感等位基因mgm101-1的ρ(+)菌株中线粒体DNA拷贝数的定量分析表明,在转移到限制温度后,每个细胞分裂时线粒体DNA的量减半。这些数据表明,在缺乏MGM101的细胞中线粒体DNA复制被迅速阻断。然而,一小部分MGM101被破坏的减数分裂分离株具有稳定维持的ρ(-)基因组。有趣的是,所有存活的ρ(-)线粒体DNA都包含一个ori/rep序列。在超抑制性(HS)菌株中破坏MGM101对HS ρ(-)线粒体DNA的增殖没有显著影响。然而,在缺乏ori/rep的小菌落中,破坏MGM101会导致线粒体DNA完全丢失或稳定性显著降低。MGM101的这种区分作用表明,ρ(+)和缺乏ori/rep的ρ(-)线粒体DNA的复制是通过相同的过程进行的。相比之下,在缺乏MGM101的HS小菌落中含有ori/rep的线粒体DNA的持续存在确定了一种独特的复制途径。ori/rep提供的替代性线粒体DNA复制机制独立于由RPO41编码的线粒体RNA聚合酶,因为HS ρ(-)基因组在mgm101、rpo41双突变体中稳定维持。