Demeshkina N A, Stiazhkina V A, Bulygin K N, Repkova M N, Ven'iaminova A G, Karpova G G
Bioorg Khim. 2005 May-Jun;31(3):295-302. doi: 10.1007/s11171-005-0036-2.
The 18S rRNA nucleotides close to the 80S ribosome template nucleotide adjacent to the A-site codon on the 3-end (i.e., the nucleotide in position +7 relative to the first nucleotide of the P-site codon) were identified using template-controlled chemical affinity ligation. For this purpose, used the photoreactive mRNA analogues with a perfluorophenylazido group attached through various linkers to the uridine C5,3'-terminal phosphate, or guanosine N7 were used. The position of the mRNA analogues on the ribosome was preset using tRNAPhe, which recognized the phenylalanine codon directed to the P-site. An analysis of the rRNAs isolated from the irradiated complexes of 80S ribosomes showed that all the analogues are almost equally ligated to the 18S rRNA nucleotides we attributed to the A-site codon environment: namely, to nucleotides A1823, A1824, and A1825 of the 3'-minidomain and to the 620-630 fragment of the 18S rRNA 5'-domain. In addition, we identified a new component of the mRNA binding site of human ribosomes, nucleotide C1698 belonging to the 18S rRNA 3-minidomain, using analogues bearing a perfluorophenylazido group on uridine and guanine residues. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 3; see also http://www.maik.ru.
利用模板控制的化学亲和连接法，鉴定了18S rRNA中靠近80S核糖体模板核苷酸的区域，该模板核苷酸位于3'端A位点密码子相邻位置（即相对于P位点密码子第一个核苷酸的+7位核苷酸）。为此，使用了光反应性mRNA类似物，这些类似物通过各种连接子连接到尿苷C5、3'-末端磷酸基团或鸟苷N7上。通过识别导向P位点的苯丙氨酸密码子的苯丙氨酰tRNA预设mRNA类似物在核糖体上的位置。对从80S核糖体的辐照复合物中分离出的rRNA进行分析表明，所有类似物几乎均等地连接到我们认为属于A位点密码子环境的18S rRNA核苷酸上，即3'-小结构域的核苷酸A1823、A1824和A1825以及18S rRNA 5'-结构域的620 - 630片段。此外，我们利用在尿苷和鸟嘌呤残基上带有全氟苯基叠氮基团的类似物，鉴定出人类核糖体mRNA结合位点的一个新组分，即属于18S rRNA 3'-小结构域的核苷酸C1698。论文英文版：《俄罗斯生物有机化学杂志》，2005年，第31卷，第3期；另见http://www.maik.ru 。
