Effect of spermine, ethylenediamine tetraacetate, urea, and high salt on the activity of ribonuclease SPL on Mg2+- containing undenatured total E. coli and rat liver RNAs.
Using RNAase SPL, a recently described nuclease from bovine seminal plasma, that can recognize certain Mg2+-dependent structural features in native naturally occurring RNAs, it is shown that these features are wholly or partially destroyed by 8 M urea, 1 mM spermine, 0.35 M KCl or NaCl, and 10 mM EDTA, urea and spermine being the most effective and EDTA the least; these features are not destroyed by heating and rapid cooling unless a medium with a high ionic strength is used. Since the buffer used so far for reconstitution of ribosomes contains 0.35 M KCl, it seems possible that a partial opening up of ribosomal RNAs may be necessary for ribosome assembly.
D000818:Animals; D004492:Edetic Acid / Q000494:pharmacology*; D004722:Endoribonucleases / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D008274:Magnesium / Q000302:isolation & purification; D009691:Nucleic Acid Denaturation; D009994:Osmolar Concentration; D012313:RNA / Q000378:metabolism*; D012329:RNA, Bacterial / Q000378:metabolism; D051381:Rats; D013096:Spermine / Q000494:pharmacology*; D014508:Urea / Q000494:pharmacology*
D012329:RNA, Bacterial; D013096:Spermine; D012313:RNA; D014508:Urea; D004492:Edetic Acid; D004722:Endoribonucleases; C032523:ribonuclease SPL; D008274:Magnesium