Zorbas Christiane, Nicolas Emilien, Wacheul Ludivine, Huvelle Emmeline, Heurgué-Hamard Valérie, Lafontaine Denis L J
RNA Molecular Biology, Fonds de la Recherche Scientifique (FRS/FNRS), Université Libre de Bruxelles, B-6041 Charleroi-Gosselies, Belgium.
Centre National de la Recherche Scientifique FRE3630, Institut de Biologie Physico-Chimique, Paris F-75005, France.
Mol Biol Cell. 2015 Jun 1;26(11):2080-95. doi: 10.1091/mbc.E15-02-0073. Epub 2015 Apr 7.
At the heart of the ribosome lie rRNAs, whose catalytic function in translation is subtly modulated by posttranscriptional modifications. In the small ribosomal subunit of budding yeast, on the 18S rRNA, two adjacent adenosines (A1781/A1782) are N(6)-dimethylated by Dim1 near the decoding site, and one guanosine (G1575) is N(7)-methylated by Bud23-Trm112 at a ridge between the P- and E-site tRNAs. Here we establish human DIMT1L and WBSCR22-TRMT112 as the functional homologues of yeast Dim1 and Bud23-Trm112. We report that these enzymes are required for distinct pre-rRNA processing reactions leading to synthesis of 18S rRNA, and we demonstrate that in human cells, as in budding yeast, ribosome biogenesis requires the presence of the modification enzyme rather than its RNA-modifying catalytic activity. We conclude that a quality control mechanism has been conserved from yeast to human by which binding of a methyltransferase to nascent pre-rRNAs is a prerequisite to processing, so that all cleaved RNAs are committed to faithful modification. We further report that 18S rRNA dimethylation is nuclear in human cells, in contrast to yeast, where it is cytoplasmic. Yeast and human ribosome biogenesis thus have both conserved and distinctive features.
核糖体的核心是核糖体RNA(rRNA),其在翻译过程中的催化功能受到转录后修饰的精细调控。在出芽酵母的小核糖体亚基中,18S rRNA上两个相邻的腺苷(A1781/A1782)在解码位点附近被Dim1进行N(6)-二甲基化,一个鸟苷(G1575)在P位点和E位点tRNA之间的一个脊处被Bud23-Trm112进行N(7)-甲基化。在这里,我们确定人类的DIMT1L和WBSCR22-TRMT112是酵母Dim1和Bud23-Trm112的功能同源物。我们报告称,这些酶对于导致18S rRNA合成的不同前体rRNA加工反应是必需的,并且我们证明,在人类细胞中,如同在出芽酵母中一样,核糖体生物发生需要修饰酶的存在而非其RNA修饰催化活性。我们得出结论,从酵母到人类,一种质量控制机制得以保留,通过该机制,甲基转移酶与新生前体rRNA的结合是加工的先决条件,以便所有切割后的RNA都能进行可靠的修饰。我们还进一步报告称,与酵母中18S rRNA二甲基化发生在细胞质中不同,在人类细胞中它发生在细胞核中。因此,酵母和人类的核糖体生物发生既有保守特征也有独特特征。