Sergiev Petr V, Serebryakova Marina V, Bogdanov Alexey A, Dontsova Olga A
Department of Chemistry and A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, 119992, Russia.
J Mol Biol. 2008 Jan 4;375(1):291-300. doi: 10.1016/j.jmb.2007.10.051. Epub 2007 Oct 23.
N(6)-Methyladenosine 1618 of Escherichia coli 23 S rRNA is located in a cluster of modified nucleotides 12 A away from the nascent peptide tunnel of the ribosome. Here, we describe the identification of gene ybiN encoding an enzyme responsible for methylation of A1618. Knockout of the ybiN gene leads to loss of modification at A1618. The modification is restored if ybiN knock-out strain has been co-transformed with a plasmid expressing the ybiN gene. On the basis of these results we suggest that ybiN gene should be renamed to rlmF in accordance with the accepted nomenclature for rRNA methyltransferases. Recombinant YbiN protein is able to methylate partially deproteinized 50 S ribosomal subunit, so-called 3.5 M LiCl core particle in vitro, but neither the completely assembled 50 S subunits nor completely deproteinized 23 S rRNA. Both lack of the ybiN gene and it's over-expression leads to growth retardation and loss of cell fitness comparative to the parental strain. It might be suggested that A1618 modification could be necessary for the exit tunnel interaction with some unknown regulatory peptides.
大肠杆菌23 S rRNA的N(6)-甲基腺苷1618位于核糖体新生肽通道12个A处的一组修饰核苷酸中。在此,我们描述了编码负责A1618甲基化的酶的基因ybiN的鉴定。ybiN基因的敲除导致A1618处修饰的缺失。如果ybiN敲除菌株与表达ybiN基因的质粒共转化,则修饰得以恢复。基于这些结果,我们建议根据rRNA甲基转移酶的公认命名法将ybiN基因重命名为rlmF。重组YbiN蛋白能够在体外使部分脱蛋白的50 S核糖体亚基(即所谓的3.5 M LiCl核心颗粒)甲基化,但不能使完全组装的50 S亚基或完全脱蛋白的23 S rRNA甲基化。与亲本菌株相比,ybiN基因的缺失及其过表达均导致生长迟缓及细胞适应性丧失。可能表明A1618修饰对于出口通道与某些未知调节肽的相互作用可能是必要的。