Nucleotides of 18S rRNA surrounding mRNA at the decoding site of translating human ribosome as revealed from the cross-linking data.

mRNA analogs, 4-(N-2-chloroethyl-N-methylamino)benzylmethyl-[5'-32P]-phosphamide derivatives of oligoribonucleotides pAUGUn (n=0, 3 or 6), were used for affinity labelling of human 80S ribosomes in complexes with codon-anticodon interaction at the P-site. These complexes were obtained in the presence of fractionated lysate from rabbit reticulocytes deprived of endogenous ribosomes and mRNAs. In all cases, 40S subunits were labelled preferentially. Within the subunits, both ribosomal proteins and 18S rRNA were modified. Ribosomal proteins cross-linked to pAUGUn derivatives were identified earlier. In this paper, nucleotides G-1010, G-1029, G-1033, G-1051, G-1054 and G-1059 of 18S rRNA cross-linked to both pAUG and pAUGU3 derivatives were identified by reverse transcription analysis. Copyright 1998 Elsevier Science B.V.