Physical properties of ribosomal ribonucleic acid isolated from bacteria deficient in ribonuclease I

Abstract

1. Bacteria deficient in ribonuclease I were used as a source of stable ribosomal RNA. RNA was isolated from a ribosome fraction of Pseudomonas fluorescens N.C.I.B. 8248 and acetone-treated cells of Escherichia coli M.R.E. 600 by the method developed by Robinson & Wade (1968). 2. The s(20,w) of the 16S and 23S components can vary from 21S and 28S down to 4S depending on the RNA macro-ion concentration and the extent to which charge is suppressed by univalent Na(+) and tris(+) counter-ions or neutralized through the binding of bivalent Mg(2+) to phosphate groups. 3. The primary charge effect in sedimentation and the frictional coefficient (which increases as the molecular conformation expands) both increase with charge and cause a decrease in s value. 4. RNA solutions heated to 80 degrees C for 10min show minor changes in s value and a detectable increase in polydispersity. Millimolar concentrations of Mg(2+) promote heat-instability and so does treatment of RNA solutions with the nuclease adsorbent macaloid, which was found to contaminate the solutions with Mg(2+). 5. The stabilization of secondary structure by univalent and bivalent cations was investigated by optical methods. 6. The sedimentation properties of 30S and 50S ribosomal subunits and their constituent 16S and 23S RNA components were compared and discussed from the viewpoint of unfolding.