Loss of ppr3, ppr4, ppr6 or ppr10 perturbs iron homeostasis and leads to apoptotic cell death in Schizosaccharomyces pombe
Loss of ppr3, ppr4, ppr6 or ppr10 perturbs iron homeostasis and leads to apoptotic cell death in Schizosaccharomyces pombe
复制标题
ppr3、ppr4、ppr6 或 ppr10 的缺失会扰乱粟酒裂殖酵母中的铁稳态并导致细胞凋亡
DOI：
10.1111/febs.13978
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Ying Huang
中科院分区：
文献类型：
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作者：
Yang Su;Yanmei Yang;Ying Huang
五肽重复序列（PPR）蛋白是一类RNA结合蛋白，参与细胞器基因表达的转录后调控。PPR蛋白广泛存在于真核生物中，在高等植物中尤为普遍，粟酒裂殖酵母（Schizoskillomycespombe）含有10种PPR蛋白。其中，ppr 3、ppr 4、ppr 6和ppr 10参与线粒体转录后过程，是线粒体电子传递链（ETC）功能所必需的。在目前的工作中，我们发现ppr 3，ppr 4，ppr 6或ppr 10的缺失导致凋亡细胞死亡，如DAPI和Annexin V-FITC染色所示。这些突变体还表现出活性氧（ROS）水平升高。RNA测序（RNA-seq）和定量RT-PCR分析显示，ppr 10的缺失影响了关键的生物学过程。特别是，参与铁摄取和/或铁稳态的核心基因组在Δ ppr 10突变体中升高，表明突变体中细胞内铁水平升高。与此一致，Δ ppr 3、Δ ppr 4、Δ ppr 6和Δ ppr 10突变体对铁的敏感性增加。此外，铁螯合剂红菲咯啉二磺酸，而不是钙螯合剂EGTA，几乎恢复了Δ ppr 3，Δ ppr 4，Δ ppr 6和Δ ppr 10突变体的活力，并降低了突变体中的ROS水平。这些结果首次表明apprgene的缺失导致铁稳态的扰动。我们的研究结果还表明，在Δ ppr 3，Δ ppr 4，Δ ppr 6，和Δ ppr 10突变体可能导致粟酒裂殖酵母中ROS水平增加和诱导细胞凋亡。数据库RNA-seq数据已保存在美国国家生物技术信息中心（NCBI）BioProject数据库中（登录号SRP 091623）和基因表达综合（GEO）数据库（登录号GSE 90144）。
Pentatricopeptide repeat (PPR) proteins characterized by tandem arrays of a degenerate 35‐amino‐acid repeat belong to a large family of RNA‐binding proteins that are involved in post‐transcriptional control of organelle gene expression. PPR proteins are ubiquitous in eukaryotes, and particularly prevalent in higher plants.Schizosaccharomyces pombehas 10 PPR proteins. Among them,ppr3,ppr4,ppr6, andppr10participate in mitochondrial post‐transcriptional processes and are required for mitochondrial electron transport chain (ETC) function. In the present work, we showed that deletion ofppr3,ppr4,ppr6, orppr10led to apoptotic cell death, as revealed by DAPI and Annexin V‐FITC staining. These mutants also exhibited elevated levels of reactive oxygen species (ROS). RNA sequencing (RNA‐seq) and quantitative RT‐PCR analyses revealed that deletion ofppr10affected critical biological processes. In particular, a core set of genes involved in iron uptake and/or iron homeostasis was elevated in the Δppr10mutant, suggesting an elevated level of intracellular iron in the mutant. Consistent with this notion, Δppr3, Δppr4, Δppr6, and Δppr10mutants exhibited increased sensitivity to iron. Furthermore, the iron chelator, bathophenanthroline disulfonic acid, but not the calcium chelator EGTA, nearly restored the viabilities of Δppr3, Δppr4, Δppr6, and Δppr10mutants, and reduced ROS levels in the mutants. These results show for the first time that deletion of apprgene leads to perturbation of iron homeostasis. Our results also suggest that disrupted iron homeostasis in Δppr3, Δppr4, Δppr6, and Δppr10mutants may lead to an increase in the level of ROS and induction of apoptotic cell death inS. pombe.DatabaseThe RNA‐seq data have been deposited in the National Center for Biotechnology Information (NCBI) BioProject database (accession number SRP091623) and Gene Expression Omnibus (GEO) database (accession number GSE90144).