Arrangement of the template 3′ of the A-site codon on the human 80S ribosome

被引：0
作者：
Molotkov M.V. [1 ]
Graifer D.M. [1 ]
Demeshkina N.A. [1 ]
Repkova M.N. [1 ]
Ven'yaminova A.G. [1 ]
Karpova G.G. [1 ]
机构：
[1] Institute of Chemical Biology and Fundamental Medicine, Siberian Division, Russian Academy of Sciences, Novosibirsk
来源：
Molecular Biology | 2005年 / 39卷 / 6期
基金：
俄罗斯基础研究基金会;
关键词：
80S ribosome; Eukaryotes; mRNA analog; Photocrosslinks; Ribosomal protein S15; rRNA;
D O I：
10.1007/s11008-005-0108-7
中图分类号：
学科分类号：
摘要：
The arrangement of the template sequence 3′ of the A-site codon on the 80S ribosome was studied using mRNA analogs containing Phe codon UUU at the 5′ end and a photoreactive perfluoroarylazido group linked to C5 of U or N7 of G. The analogs were positioned on the ribosome with the use of tRNA Phe, which directed the UUU codon to the P site, bringing a modified nucleotide to position +9 or +12 relative to the first nucleotide of the P-site codon. Upon mild UV irradiation of ribosome complexes, the analogs of both types crosslinked to the 18S rRNA and proteins of the 40S subunit. Comparisons were made with the crosslinking patterns of complexes in which an mRNA analog contained a modified nucleotide in position +7 (the crosslinking to 18S rRNA in such complexes has been studied previously). The efficiency of crosslinking to ribosomal components depended on the nature of the modified nucleotide of an mRNA analog and its position on the ribosome. The extent of crosslinking to the 18S rRNA drastically decreased as the modified nucleotide was transferred from position +7 to position +12. The 18S rRNA nucleotides involved in crosslinking were identified. A modified nucleotide in position +9 crosslinked to the invariant dinucleotide A1824/A1825 and variable A1823 in the 3′ minidomain of the 18S rRNA and to S15. The same ribosomal components have earlier been shown to crosslink to modified nucleotides in positions +4 to +7. In addition, all mRNA analogs crosslinked to invariant C1698 in the 3′ minidomain and to conserved region 605-620, which closes helix 18 in the 5′ domain. © 2005 Pleiades Publishing, Inc.
引用
收藏
页码：876 / 883
页数：7
相关论文
共 29 条
  • [1] Sergiev P.V., Dontsova O.A., Bogdanov A.A., Studying the structure of the prokaryotic ribosome by biochemical methods: The Judgment Day, Mol. Biol., 35, pp. 559-583, (2001)
  • [2] Yusupova G.Zh., Yusupov M.M., Cate J.H.D., Noller H.F., The path of messenger RNA through the ribosome, Cell., 106, pp. 233-241, (2001)
  • [3] Ogle J.M., Brodersen D.E., Clemons W.M., Tarry M.J., Carter A.P., Ramakrishnan V., Recognition of cognate transfer RNA by the 30S ribosomal subunit, Science, 292, pp. 897-902, (2001)
  • [4] Ramakrishnan V., Ribosome structure and the mechanism of translation, Cell., 108, pp. 557-572, (2002)
  • [5] Graifer D.M., Karpova G.G., Structutal-functional topography of human ribosomes as inferred from data on crosslinking with oligoribonucleotide-derived mRNA analogs, Mol. Biol., 35, pp. 584-596, (2001)
  • [6] Demeshkina N., Repkova M., Ven'yaminova A., Graifer D., Karpova G., Nucleotides of 18S rRNA surrounding mRNA codons at the human ribosomal A, P and E sites, respectively: A crosslinking study with mRNA analogues carrying aryl azide group at either the uracil or the guanine residue, RNA, 6, pp. 1727-1736, (2000)
  • [7] Demeshkina N., Laletina E., Meschaninova M., Ven'yaminova A., Graifer D., Karpova G., Positioning of mRNA codons with respect to 18S rRNA at the P and E sites of human ribosome, Biochim. Biophys. Acta, 1627, pp. 39-46, (2003)
  • [8] Graifer D., Molotkov M., Styazhkina V., Demeshkina N., Bulygin K., Eremina A., Ivanov A., Laletina E., Ven'yaminova A., Karpova G., Variable and conserved elements of human ribosomes surrounding the mRNA at the decoding and upstream sites, Nucleic Acids Res., 32, pp. 3282-3293, (2004)
  • [9] Spann C.M.T., Beckmann R., Eswar N., Penczek P.A., Sali A., Blobel G., Frank J., Structure of the 80S ribosome from Saccaromyces cerevisiae: tRNA-ribosome and subunit-subunit interactions, Cell., 107, pp. 373-386, (2001)
  • [10] Morgan D.G., Menetret J.-F., Neuhof A., Rapoport T.A., Akey C.W., Structure of the mammalian ribosome-channel complex at 17 Åresolution, J. Mol. Biol., 324, pp. 871-886, (2002)
123