Resistance of both the 5'- and 3'-domain of isolated Escherichia coli 23S rRNA against digestion with alpha-sarcin.
The ribonuclease alpha-sarcin exclusively cleaves the phosphodiester bond after G2661 in the 23S rRNA within 50S subunits, thus inactivating the ribosomes. The resulting alpha-fragment is 243 nucleotides long and contains the 3'-end of the 23S rRNA. The specificity is changed dramatically if isolated 23S rRNA is used as substrate. We have shown previously that 23S rRNA is digested completely except for two fragments, one of which is identical to the alpha-fragment. Here we show that the other fragment comprises the 5'-end of 23S rRNA and contains 385 nucleotides. A similar fragment was obtained when isolated 23S rRNA was digested with RNase A (specific for pyrimidines in single strands). It appears that the 5'-domain (equivalent to 5.8S rRNA of eukaryotic ribosomes) as well as the 3'-domain (equivalent to 4.5S rRNA of chloroplast ribosomes) have a compact and defined tertiary structure in isolated 23S rRNA in contrast to the rRNA region in between. Thus, alpha-sarcin is a convenient tool for detecting compact domains in isolated RNA.
Hausner
T P
TP
Max-Planck-Institut für Molekulare Genetik, Abteilung Wittman, Berlin-Dahlem.
Nierhaus
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KH
D001483:Base Sequence; D004722:Endoribonucleases; D004926:Escherichia coli / Q000235:genetics*; D005656:Fungal Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D011500:Protein Synthesis Inhibitors / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000378:metabolism*; D012338:RNA, Ribosomal, 23S / Q000378:metabolism*; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D013379:Substrate Specificity
D005656:Fungal Proteins; D011500:Protein Synthesis Inhibitors; D012335:RNA, Ribosomal; D012338:RNA, Ribosomal, 23S; C014801:alpha-sarcin; D004722:Endoribonucleases; D012259:Ribonuclease, Pancreatic