Department of Systems Biotechnology, Chung-Ang University, Anseong, 17546, Republic of Korea.
School of Biological Sciences, Seoul National University, Seoul, 08826, Republic of Korea.
J Microbiol. 2021 Dec;59(12):1075-1082. doi: 10.1007/s12275-021-1438-4. Epub 2021 Oct 26.
Aconitase, a highly conserved protein across all domains of life, functions in converting citrate to isocitrate in the tricarboxylic acid cycle. Cytosolic aconitase is also known to act as an iron regulatory protein in mammals, binding to the RNA hairpin structures known as iron-responsive elements within the untranslated regions of specific RNAs. Aconitase-2 (Aco2) in fission yeast is a fusion protein consisting of an aconitase and a mitochondrial ribosomal protein, bL21, residing not only in mitochondria but also in cytosol and the nucleus. To investigate the role of Aco2 in the nucleus and cytoplasm of fission yeast, we analyzed the transcriptome of aco2ΔN mutant that is deleted of nuclear localization signal (NLS). RNA sequencing revealed that the aco2ΔN mutation caused increase in mRNAs encoding iron uptake transporters, such as Str1, Str3, and Shu1. The half-lives of mRNAs for these genes were found to be significantly longer in the aco2ΔN mutant than the wild-type strain, suggesting the role of Aco2 in mRNA turnover. The three conserved cysteines required for the catalytic activity of aconitase were not necessary for this role. The UV cross-linking RNA immunoprecipitation analysis revealed that Aco2 directly bound to the mRNAs of iron uptake transporters. Aco2-mediated degradation of iron-uptake mRNAs appears to utilize exoribonuclease pathway that involves Rrp6 as evidenced by genetic interactions. These results reveal a novel role of non-mitochondrial aconitase protein in the mRNA turnover in fission yeast to fine-tune iron homeostasis, independent of regulation by transcriptional repressor Fep1.
aconitase,一种在所有生命领域都高度保守的蛋白质,在三羧酸循环中催化柠檬酸转化为异柠檬酸。细胞质 aconitase 也被认为是哺乳动物中的一种铁调节蛋白,与未翻译区域中特定 RNA 的 RNA 发夹结构(称为铁反应元件)结合。裂殖酵母中的 aconitase-2 (Aco2) 是一种融合蛋白,由 aconitase 和线粒体核糖体蛋白 bL21 组成,不仅存在于线粒体中,还存在于细胞质和核中。为了研究 Aco2 在裂殖酵母核和细胞质中的作用,我们分析了缺失核定位信号(NLS)的 aco2ΔN 突变体的转录组。RNA 测序表明,aco2ΔN 突变导致编码铁摄取转运体的 mRNA 增加,如 Str1、Str3 和 Shu1。发现这些基因的 mRNA 半衰期在 aco2ΔN 突变体中比野生型菌株显著延长,表明 Aco2 在 mRNA 周转中起作用。 aconitase 催化活性所需的三个保守半胱氨酸对于这种作用不是必需的。UV 交联 RNA 免疫沉淀分析表明,Aco2 直接与铁摄取转运体的 mRNA 结合。Aco2 介导的铁摄取 mRNA 降解似乎利用了涉及 Rrp6 的外切核酸酶途径,这一点可以从遗传相互作用中得到证明。这些结果揭示了非线粒体 aconitase 蛋白在裂殖酵母 mRNA 周转中的新作用,以微调铁稳态,独立于转录抑制剂 Fep1 的调节。