线粒体 DNA 与 RNA 之间的异质体一致性。

Heteroplasmy concordance between mitochondrial DNA and RNA.

机构信息

Division of Nutritional Sciences, Cornell University, Ithaca, New York, 14853, USA.

Regeneron Pharmaceuticals, Inc, Tarrytown, NY, 10591, USA.

出版信息

Sci Rep. 2019 Sep 10;9(1):12942. doi: 10.1038/s41598-019-49279-7.

Abstract

Mitochondrial DNA (mtDNA) heteroplasmies are associated with various diseases but the transmission of heteroplasmy from mtDNA to mitochondrial RNA (mtRNA) remains unclear. We compared heteroplasmies in mtRNA from 446 human B-lymphoblastoid cell lines to their corresponding mtDNA using deep sequencing data from two independent studies. We observed 2786 heteroplasmies presenting in both DNA and RNA at 1% frequency cutoff. Among them, the frequencies of 2427 (87.1%) heteroplasmies were highly consistent (less than 5% frequency difference) between DNA and RNA. To validate these frequency consistencies, we isolated DNA and RNA simultaneously from GM12282 cell line used in those two sequencing studies, and resequenced its heteroplasmy sites. Interestingly, we also observed the rapid changes of heteroplasmy frequencies during 4 weeks of the cell culture: the frequencies at Day 14 increased by >25% than those at Day 0. However, the heteroplasmy frequencies from the same time point were highly consistent. In summary, our analysis on public data together with in vitro study indicates that the heteroplasmies in DNA can be transcribed into RNA with high fidelity. Meanwhile, the observed rapid-changing heteroplasmy frequency can potentially disturb cell functions, which could be an overlooked confounding factor in cell line related studies.

摘要

线粒体 DNA(mtDNA)异质性与多种疾病有关,但 mtDNA 到线粒体 RNA(mtRNA)的异质性传递仍不清楚。我们使用来自两项独立研究的深度测序数据,比较了 446 个人类 B 淋巴细胞系的 mtRNA 中的异质性与其相应的 mtDNA。我们观察到在 DNA 和 RNA 中以 1%频率截止值呈现的 2786 种异质性。其中,2427 种(87.1%)异质性的频率在 DNA 和 RNA 之间高度一致(频率差异小于 5%)。为了验证这些频率一致性,我们从用于这两项测序研究的 GM12282 细胞系中同时分离 DNA 和 RNA,并重新测序其异质性位点。有趣的是,我们还观察到细胞培养期间异质性频率的快速变化:第 14 天的频率比第 0 天增加了>25%。然而,同一时间点的异质性频率高度一致。总之,我们对公共数据的分析和体外研究表明,DNA 中的异质性可以以高保真度转录为 RNA。同时,观察到的快速变化的异质性频率可能会干扰细胞功能,这可能是细胞系相关研究中被忽视的混杂因素。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8925/6737107/b98ebc2774f2/41598_2019_49279_Fig1_HTML.jpg

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