Yarmishyn Aliaksandr A, Kremenskoy Maksym, Batagov Arsen O, Preuss Axel, Wong Jin Huei, Kurochkin Igor V
Department of Genome and Gene Expression Data Analysis, Bioinformatics Institute, Agency for Science, Technology and Research (A*STAR), Matrix, Singapore, 138671, Singapore.
Institute of Molecular and Cellular Biology, Agency for Science, Technology and Research (A*STAR), Proteos, Singapore, 138673, Singapore.
BMC Genomics. 2016 Dec 22;17(Suppl 13):1028. doi: 10.1186/s12864-016-3330-x.
RNA is often targeted to be localized to the specific subcellular compartments. Specific localization of mRNA is believed to be an important mechanism for targeting their protein products to the locations, where their function is required.
In this study we performed the genome wide transcriptome analysis of peroxisome preparations from the mouse liver using microarrays. We demonstrate that RNA is absent inside peroxisomes, however it is associated at their exterior via the noncovalent contacts with the membrane proteins. We detect enrichment of specific sets of transcripts in two preparations of peroxisomes, purified with different degrees of stringency. Importantly, among these were mRNAs encoding bona fide peroxisomal proteins, such as peroxins and peroxisomal matrix enzymes involved in beta-oxidation of fatty acids and bile acid biosynthesis. The top-most enriched mRNA, whose association with peroxisomes we confirm microscopically was Hmgcs1, encoding 3-hydroxy-3-methylglutaryl-CoA synthase, a crucial enzyme of cholesterol biosynthesis pathway. We observed significant representation of mRNAs encoding mitochondrial and secreted proteins in the peroxisomal fractions.
This is a pioneer genome-wide study of localization of mRNAs to peroxisomes that provides foundation for more detailed dissection of mechanisms of RNA targeting to subcellular compartments.
RNA常常被靶向定位于特定的亚细胞区室。信使核糖核酸（mRNA）的特异性定位被认为是将其蛋白质产物靶向至需要其功能的位置的重要机制。
在本研究中，我们使用微阵列对来自小鼠肝脏的过氧化物酶体提取物进行了全基因组转录组分析。我们证明RNA不存在于过氧化物酶体内部，然而它通过与膜蛋白的非共价接触而与过氧化物酶体外部相关联。我们在两种以不同严格程度纯化的过氧化物酶体制剂中检测到特定转录本组的富集。重要的是，其中包括编码真正过氧化物酶体蛋白的mRNA，例如参与脂肪酸β氧化和胆汁酸生物合成的过氧化物酶和过氧化物酶体基质酶。我们通过显微镜确认与过氧化物酶体相关联的最富集的mRNA是Hmgcs1，它编码3-羟基-3-甲基戊二酰辅酶A合酶，这是胆固醇生物合成途径中的一种关键酶。我们在过氧化物酶体组分中观察到编码线粒体和分泌蛋白的mRNA有显著表现。
这是一项关于mRNA定位于过氧化物酶体的全基因组开创性研究，为更详细剖析RNA靶向亚细胞区室的机制奠定了基础。
