Differential regulation of full-length genome and a single-stranded 7S DNA along the cell cycle in human mitochondria.

Differential regulation of full-length genome and a single-stranded 7S DNA along the cell cycle in human mitochondria.
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DOI:
10.1093/nar/gkq493
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发表时间:
2010-10
影响因子:
14.9
通讯作者:
Lee CG
Lee CG
中科院分区:
生物学2区
文献类型:
--
作者:
Antes A;Tappin I;Chung S;Lim R;Lu B;Parrott AM;Hill HZ;Suzuki CK;Lee CG

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哺乳动物线粒体含有全长基因组和单链 7S DNA。尽管线粒体 DNA (mtDNA) 的拷贝数根据细胞类型以及对不同环境压力的反应而变化,但我们对 mtDNA 和 7S DNA 如何维持和调节的理解有限,部分原因是缺乏反映线粒体体内功能的可靠体外测定系统。在这里,我们报告了一种体外测定系统,用于在可控的体外条件下测量 mtDNA 和 7S DNA 的合成。通过该检测系统,我们证明线粒体的复制能力与 mtDNA 和 7S DNA 的内源拷贝数相关。我们的研究还表明,较高的核苷酸浓度会越来越促进 7S DNA 合成,但不会促进 mtDNA 合成。一致的是,线粒体合成 7S DNA 但不合成 mtDNA 的能力在细胞周期中发生显着变化,在 S 期达到最高水平。这些发现表明,mtDNA 和 7S DNA 的合成是独立进行的,并且线粒体合成 7S DNA 的能力不仅随着细胞周期的进展而动态变化,而且还响应不同的核苷酸浓度而动态变化。
Mammalian mitochondria contain full-length genome and a single-stranded 7S DNA. Although the copy number of mitochondrial DNA (mtDNA) varies depending on the cell type and also in response to diverse environmental stresses, our understanding of how mtDNA and 7S DNA are maintained and regulated is limited, partly due to lack of reliable in vitro assay systems that reflect the in vivo functionality of mitochondria. Here we report an in vitro assay system to measure synthesis of both mtDNA and 7S DNA under a controllable in vitro condition. With this assay system, we demonstrate that the replication capacity of mitochondria correlates with endogenous copy numbers of mtDNA and 7S DNA. Our study also shows that higher nucleotide concentrations increasingly promote 7S DNA synthesis but not mtDNA synthesis. Consistently, the mitochondrial capacity to synthesize 7S DNA but not mtDNA noticeably varied along the cell cycle, reaching its highest level in S phase. These findings suggest that syntheses of mtDNA and 7S DNA proceed independently and that the mitochondrial capacity to synthesize 7S DNA dynamically changes not only with cell-cycle progression but also in response to varying nucleotide concentrations.
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