[Template location on the human ribosome: environment of the mRNA nucleotide adjacent to the A-site codon on the 3'-side].
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.
Demeshkina
N A
NA
Stiazhkina
V A
VA
Bulygin
K N
KN
Repkova
M N
MN
Ven'iaminova
A G
AG
Karpova
G G
GG
D003062:Codon / Q000737:chemistry*; D006801:Humans; D012333:RNA, Messenger / Q000737:chemistry*; D012337:RNA, Ribosomal, 18S / Q000737:chemistry*; D012360:RNA, Transfer, Phe / Q000737:chemistry; D012265:Ribonucleotides / Q000737:chemistry*; D012270:Ribosomes / Q000737:chemistry*
D003062:Codon; D012333:RNA, Messenger; D012337:RNA, Ribosomal, 18S; D012360:RNA, Transfer, Phe; D012265:Ribonucleotides