PNPase knockout results in mtDNA loss and an altered metabolic gene expression program.
PNPase knockout results in mtDNA loss and an altered metabolic gene expression program.
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DOI：
10.1371/journal.pone.0200925
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Teitell MA
中科院分区：
文献类型：
--
作者：
Shimada E;Ahsan FM;Nili M;Huang D;Atamdede S;TeSlaa T;Case D;Yu X;Gregory BD;Perrin BJ;Koehler CM;Teitell MA
多核苷酸磷酸化酶（Polynucleotide phosphorylase，PNIPs）是一种重要的核糖核酸外切酶，参与多种生物学过程和人类疾病。为了揭示PNTR在线粒体中的作用，我们通过首先改变培养条件以使具有缺陷呼吸的细胞生长来建立PNTR敲除（PKO）系统。有趣的是，在小鼠胚胎成纤维细胞（MEFs）中建立的PKO导致线粒体DNA（mtDNA）的丢失。PKO细胞的转录谱与rho 0 mtDNA缺失细胞相似，与野生型亲本（TM 6）MEFs相比，胆固醇（FDR = 6.35 x 10−13）、脂质（FDR = 3.21 x 10−11）和仲醇（FDR = 1.04 x 10 -12）代谢途径基因表达受到干扰。转录组分析表明，与轴突发生（FDR = 4.49 x 10−3）、轴突发育（FDR = 4.74 x 10−3）和轴突导向（FDR = 4.74 x 10 −3）相关的过程在PKO细胞中过度表达，这与先前详细描述人类髓鞘形成延迟、听力损失、脑肌病和脉络膜视网膜缺陷中的致病性PNIPs突变的研究一致。过度表达分析显示，在PKO和rho 0细胞的代谢途径的改变。因此，我们评估了与细胞周期进程和总代谢有关的基因的相关性，并观察到PKO细胞与rho 0 MEFs之间的强正相关性，而与TM 6 MEFs相比。我们定量PKO克隆的归一化生物量积累速率为1.7%（SD ± 2.0%）和2.4%（SD ± 1.6%）/小时，低于TM 6细胞的3.3%（SD ± 3.5%）/小时。此外，小鼠内耳毛细胞中的PKO导致进行性听力损失，这与先前与PNTR突变相关的人类家族性听力损失相似。结合，我们的研究报告，敲除线粒体核酸酶导致mtDNA的损失，并表明mtDNA的维护可以提供一个统一的连接，为大量的生物学活动报告的PNTR。
Polynucleotide phosphorylase (PNPase) is an essential mitochondria-localized exoribonuclease implicated in multiple biological processes and human disorders. To reveal role(s) for PNPase in mitochondria, we established PNPase knockout (PKO) systems by first shifting culture conditions to enable cell growth with defective respiration. Interestingly, PKO established in mouse embryonic fibroblasts (MEFs) resulted in the loss of mitochondrial DNA (mtDNA). The transcriptional profile of PKO cells was similar to rho0 mtDNA deleted cells, with perturbations in cholesterol (FDR = 6.35 x 10−13), lipid (FDR = 3.21 x 10−11), and secondary alcohol (FDR = 1.04x10-12) metabolic pathway gene expression compared to wild type parental (TM6) MEFs. Transcriptome analysis indicates processes related to axonogenesis (FDR = 4.49 x 10−3), axon development (FDR = 4.74 x 10−3), and axonal guidance (FDR = 4.74 x 10−3) were overrepresented in PKO cells, consistent with previous studies detailing causative PNPase mutations in delayed myelination, hearing loss, encephalomyopathy, and chorioretinal defects in humans. Overrepresentation analysis revealed alterations in metabolic pathways in both PKO and rho0 cells. Therefore, we assessed the correlation of genes implicated in cell cycle progression and total metabolism and observed a strong positive correlation between PKO cells and rho0 MEFs compared to TM6 MEFs. We quantified the normalized biomass accumulation rate of PKO clones at 1.7% (SD ± 2.0%) and 2.4% (SD ± 1.6%) per hour, which was lower than TM6 cells at 3.3% (SD ± 3.5%) per hour. Furthermore, PKO in mouse inner ear hair cells caused progressive hearing loss that parallels human familial hearing loss previously linked to mutations in PNPase. Combined, our study reports that knockout of a mitochondrial nuclease results in mtDNA loss and suggests that mtDNA maintenance could provide a unifying connection for the large number of biological activities reported for PNPase.
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影响因子：
48
作者：
Huber W;Carey VJ;Gentleman R;Anders S;Carlson M;Carvalho BS;Bravo HC;Davis S;Gatto L;Girke T;Gottardo R;Hahne F;Hansen KD;Irizarry RA;Lawrence M;Love MI;MacDonald J;Obenchain V;Oleś AK;Pagès H;Reyes A;Shannon P;Smyth GK;Tenenbaum D;Waldron L;Morgan M
通讯作者：
Morgan M
DOI：
10.1073/pnas.0900221106
发表时间：
2009-06-16
影响因子：
11.1
作者：
Belyantseva, Inna A.;Perrin, Benjamin J.;Ervasti, James M.
通讯作者：
Ervasti, James M.
影响因子：
3.6
作者：
Goverde, RLJ;Veld, JHJHI;Mooi, FR
通讯作者：
Mooi, FR
影响因子：
5.3
作者：
Deng, Suhua;Hirschberg, Alexandra;Kuner, Rohini
通讯作者：
Kuner, Rohini
影响因子：
56.9
作者：
GRUNBERGMANAGO, M;ORTIZ, PJ;OCHOA, S
通讯作者：
OCHOA, S