Zhan Yu, Dhaliwal James S, Adjibade Pauline, Uniacke James, Mazroui Rachid, Zerges William
Biology Department & Centre for Structural and Functional Genomics, Concordia University, 7141 Sherbrooke W, Montreal, Quebec, Canada H4B 1R6.
Department of Molecular Biology, Medical Biochemistry, and Pathology, Laval University, Centre de Recherche le CHU de Quebec, Quebec, Canada G1V 4G2.
J Cell Sci. 2015 Nov 15;128(22):4210-9. doi: 10.1242/jcs.175232. Epub 2015 Oct 8.
The oxidation of biological molecules by reactive oxygen species (ROS) can render them inactive or toxic. This includes the oxidation of RNA, which appears to underlie the detrimental effects of oxidative stress, aging and certain neurodegenerative diseases. Here, we investigate the management of oxidized RNA in the chloroplast of the green alga Chlamydomonas reinhardtii. Our immunofluorescence microscopy results reveal that oxidized RNA (with 8-hydroxyguanine) is localized in the pyrenoid, a chloroplast microcompartment where CO2 is assimilated by the Calvin cycle enzyme Rubisco. Results of genetic analyses support a requirement for the Rubisco large subunit (RBCL), but not Rubisco, in the management of oxidized RNA. An RBCL pool that can carry out such a 'moonlighting' function is revealed by results of biochemical fractionation experiments. We also show that human (HeLa) cells localize oxidized RNA to cytoplasmic foci that are distinct from stress granules, processing bodies and mitochondria. Our results suggest that the compartmentalization of oxidized RNA management is a general phenomenon and therefore has some fundamental significance.
活性氧(ROS)对生物分子的氧化作用会使其失去活性或产生毒性。这包括RNA的氧化,而这似乎是氧化应激、衰老及某些神经退行性疾病产生有害影响的潜在原因。在此,我们研究了绿藻莱茵衣藻叶绿体中氧化RNA的处理机制。我们的免疫荧光显微镜检查结果显示,氧化RNA(含8-羟基鸟嘌呤)定位于蛋白核中,蛋白核是叶绿体的一个微区室,二氧化碳通过卡尔文循环酶核酮糖-1,5-二磷酸羧化酶/加氧酶(Rubisco)在此处被同化。遗传分析结果支持在氧化RNA的处理过程中需要核酮糖-1,5-二磷酸羧化酶大亚基(RBCL),而非核酮糖-1,5-二磷酸羧化酶。生化分级分离实验结果揭示了一个能够执行这种“兼职”功能的RBCL库。我们还表明,人类(HeLa)细胞将氧化RNA定位于与应激颗粒、加工小体和线粒体不同的细胞质病灶中。我们的结果表明,氧化RNA处理的区室化是一种普遍现象,因此具有一些基本意义。