Variable and conserved elements of human ribosomes surrounding the mRNA at the decoding and upstream sites

被引:59
作者
Graifer, D [1 ]
Molotkov, M [1 ]
Styazhkina, V [1 ]
Demeshkina, N [1 ]
Bulygin, K [1 ]
Eremina, A [1 ]
Ivanov, A [1 ]
Laletina, E [1 ]
Ven'yaminova, A [1 ]
Karpova, G [1 ]
机构
[1] Russian Acad Sci, Siberian Branch, Inst Chem Biol & Fundamental Med, Novosibirsk 630090, Russia
基金
俄罗斯基础研究基金会;
关键词
D O I
10.1093/nar/gkh657
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
This study is centred upon an important biological problem concerning the structural organization of mammalian ribosomes that cannot be studied by X-ray analysis because 80S ribosome crystals are still unavailable. Here, positioning of the mRNA on 80S ribosomes was studied using mRNA analogues containing the perfluorophenylazide cross-linker on either the guanosine or an uridine residue. The modi-fied nucleotides were directed to positions from -9 to +6 with respect to the first nucleotide of the P site bound codon by a tRNA cognate to the triplet targeted to the P site. Upon mild UV-irradiation, the modified nucleotides at positions +4 to +6 cross-linked to protein S15 and 18S rRNA nucleotides A1823-A1825. In addition, modified guanosines in positions +5 and +6 also cross-linked to G626, and in position +1 to G1702. Cross-linking from the upstream positions was mainly to protein S26 that has no prokaryotic homologues. These findings indicate that the tail of mammalian S15 comes closer to the decoding site than that of its prokaryotic homologue S19, and that the environments of the upstream part of mRNA on 80S and 70S ribosomes differ. On the other hand, the results confirm the widely accepted idea regarding the conserved nature of the decoding site of the small subunit rRNA.
引用
收藏
页码:3282 / 3293
页数:12
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