Department of Biochemistry, Institute of Molecular and Cell Biology, University of Tartu, Tartu, Estonia.
Mol Cell. 2010 Sep 24;39(6):851-61. doi: 10.1016/j.molcel.2010.09.002.
Molecular recombination and transcription are proposed mechanisms to initiate mitochondrial DNA (mtDNA) replication in yeast. We conducted a comprehensive analysis of mtDNA from the yeast Candida albicans. Two-dimensional agarose gel electrophoresis of mtDNA intermediates reveals no bubble structures diagnostic of specific replication origins, but rather supports recombination-driven replication initiation of mtDNA in yeast. Specific species of Y structures together with DNA copy number analyses of a C. albicans mutant strain provide evidence that a region in a mainly noncoding inverted repeat is predominantly involved in replication initiation via homologous recombination. Our further findings show that the C. albicans mtDNA forms a complex branched network that does not contain detectable amounts of circular molecules. We provide topological evidence for recombination-driven mtDNA replication initiation and introduce C. albicans as a suitable model organism to study wild-type mtDNA maintenance in yeast.
分子重组和转录被提出是启动酵母中线粒体 DNA(mtDNA)复制的机制。我们对来自酵母白色念珠菌的 mtDNA 进行了全面分析。mtDNA 中间体的二维琼脂糖凝胶电泳显示没有可诊断特定复制起点的泡状结构,而是支持酵母中线粒体 DNA 的重组驱动复制起始。Y 结构的特定物种以及白色念珠菌突变株的 DNA 拷贝数分析提供了证据,表明主要是非编码反向重复中的一个区域主要通过同源重组参与复制起始。我们的进一步发现表明,白色念珠菌的 mtDNA 形成了一个复杂的分支网络,其中不包含可检测量的环状分子。我们提供了拓扑学证据,证明重组驱动的 mtDNA 复制起始,并将白色念珠菌作为研究酵母中野生型 mtDNA 维持的合适模式生物。