PMID:9149228
| Citation |
Sergiev, PV, Lavrik, IN, Wlasoff, VA, Dokudovskaya, SS, Dontsova, OA, Bogdanov, AA and Brimacombe, R (1997) The path of mRNA through the bacterial ribosome: a site-directed crosslinking study using new photoreactive derivatives of guanosine and uridine. RNA 3:464-75 |
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| Abstract |
Two new photoreactive nucleotide derivatives have been applied in site-directed crosslinking studies with mRNA analogues. 6-Thioguanosine triphosphate or 5-methyleneaminouridine triphosphate was incorporated into mRNA analogues by T7 transcription; after transcription, the 5-methyleneaminouridine residues were converted to a diazirine derivative. mRNA analogues carrying either 6-thioguanosine or the diazirine derivative were bound to Escherichia coli ribosomes in the presence of tRNA(f)(Met), and photo-crosslinking was induced by irradiation at 350 nm. With 6-thioguanosine, specific crosslinks were observed from downstream positions +8 or +9 of the mRNA to nt 1196 in helix 34 of the 16S rRNA, and from position +12 to nt 530 in helix 18. With the diazirine derivative, a crosslink from position +2 (within the AUG codon) to nt 926 in helix 28 was found. Taken together with previous data obtained from downstream sites in mRNA analogues carrying 4-thiouridine residues, specific crosslinks have now been identified from downstream mRNA positions +2, +4, +6, +7, +8, +9, +11, and +12. The data confirm that the three 16S rRNA regions involved-helices 18, 28, and 34-are in the direct neighborhood of the decoding area of the 30S subunit. |
| Links | |
| Keywords |
Bacteriophage T7/metabolism; Base Sequence; Codon; Cross-Linking Reagents; Escherichia coli/metabolism; Guanosine Triphosphate/analogs & derivatives; Guanosine Triphosphate/chemical synthesis; Guanosine Triphosphate/metabolism; Molecular Sequence Data; Nucleic Acid Conformation; RNA, Bacterial/biosynthesis; RNA, Bacterial/chemistry; RNA, Bacterial/metabolism; RNA, Messenger/biosynthesis; RNA, Messenger/chemistry; RNA, Messenger/metabolism; RNA, Ribosomal, 16S/biosynthesis; RNA, Ribosomal, 16S/chemistry; RNA, Transfer, Met/metabolism; Ribosomes/metabolism; Thionucleotides/chemical synthesis; Thionucleotides/metabolism; Transcription, Genetic; Uridine Triphosphate/analogs & derivatives; Uridine Triphosphate/chemical synthesis; Uridine Triphosphate/metabolism |
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