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Bud23 Methylates G1575 of 18S rRNA and Is Required for Efficient Nuclear Export of Pre-40S Subunits

机译:18S rRNA的Bud23甲酯G1575对于有效地输出40S之前的亚基是必需的

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摘要

BUD23 was identified from a bioinformatics analysis of Saccharomyces cerevisiae genes involved in ribosome biogenesis. Deletion of BUD23 leads to severely impaired growth, reduced levels of the small (40S) ribosomal subunit, and a block in processing 20S rRNA to 18S rRNA, a late step in 40S maturation. Bud23 belongs to the S-adenosylmethionine-dependent Rossmann-fold methyltransferase superfamily and is related to small-molecule methyltransferases. Nevertheless, we considered that Bud23 methylates rRNA. Methylation of G1575 is the only mapped modification for which the methylase has not been assigned. Here, we show that this modification is lost in bud23 mutants. The nuclear accumulation of the small-subunit reporters Rps2-green fluorescent protein (GFP) and Rps3-GFP, as well as the rRNA processing intermediate, the 5′ internal transcribed spacer 1, indicate that bud23 mutants are defective for small-subunit export. Mutations in Bud23 that inactivated its methyltransferase activity complemented a bud23Δ mutant. In addition, mutant ribosomes in which G1575 was changed to adenosine supported growth comparable to that of cells with wild-type ribosomes. Thus, Bud23 protein, but not its methyltransferase activity, is important for biogenesis and export of the 40S subunit in yeast.
机译:从涉及核糖体生物发生的啤酒酵母基因的生物信息学分析中鉴定出BUD23。缺失BUD23会导致生长严重受损,小的(40S)核糖体亚基水平降低,以及将20S rRNA加工为18S rRNA的障碍,这是40S成熟的后期步骤。 Bud23属于S-腺苷甲硫氨酸依赖性的Rossmann折叠甲基转移酶超家族,与小分子甲基转移酶有关。尽管如此,我们认为Bud23甲基化了rRNA。 G1575的甲基化是唯一未分配甲基化酶的定位图谱。在这里,我们表明这种修饰在bud23突变体中丢失。小亚基报告基因Rps2-绿色荧光蛋白(GFP)和Rps3-GFP的核积累,以及rRNA加工中间体,即5'内部转录的间隔区1,表明bud23突变体对于小亚基输出是有缺陷的。 Bud23中的甲基转移酶活性失活的突变与bud23Δ突变互补。此外,其中G1575变为腺苷的突变核糖体支持的生长与野生型核糖体的细胞相当。因此,Bud23蛋白而不是其甲基转移酶活性对于酵母中40S亚基的生物发生和输出很重要。

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