Mapping RNA regions in eukaryotic ribosomes that are accessible to methidiumpropyl-EDTA.Fe(II) and EDTA.Fe(II).
Methidiumpropyl-EDTA.Fe(II) [MPE.Fe(II)] and EDTA.Fe(II) were used to investigate the structure of Drosophila melanogaster ribosomes. Cleavage reactions were performed on intact ribosomes in cell lysates in vitro and analyzed by primer extension with reverse transcriptase using oligodeoxynucleotide primers. Regions of 18S and 28S ribosomal RNAs (rRNAs) which are accessible to MPE.Fe(II) and EDTA.Fe(II) are located almost exclusively within expansion segments. The accessibility of these regions to cleavage indicates that they are likely exposed on the surface of eukaryotic ribosomes. These results provide information about the overall tertiary structure of rRNA in ribosomes.
Han
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Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena 91125.
Schepartz
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Pellegrini
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Dervan
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D000818:Animals; D001345:Autoradiography; D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry; D004331:Drosophila melanogaster / Q000196:embryology / Q000235:genetics*; D004492:Edetic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D005296:Ferrous Compounds / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012335:RNA, Ribosomal / Q000737:chemistry*; D012337:RNA, Ribosomal, 18S / Q000737:chemistry; D012339:RNA, Ribosomal, 28S / Q000737:chemistry; D012270:Ribosomes / Q000737:chemistry* / Q000187:drug effects
D017931:DNA Primers; D005296:Ferrous Compounds; D012335:RNA, Ribosomal; D012337:RNA, Ribosomal, 18S; D012339:RNA, Ribosomal, 28S; C038200:methidiumpropyl-EDTA-iron(II); C055526:Fe(II)-EDTA; D004492:Edetic Acid