Summary
Disruption of multiplying bacteria by pressure release produces the dissociation of polysomes and the formation of ribosomal subunits, which quickly reassociate into 70S ribosomes. Most reassociated particles are stable when centrifuged at high speed in neutral sucrose gradients containing 10 mM Mg++. Several environmental factors prevent the reassociation of pressure-dissociated ribosomes: pH<6, NaN3>50mM, NH4Cl>100 mM, Mg++<6 mM, and glutaraldehyde >0.1%. When cellular growth is blocked either by nutrient exhaustion (stationary phase, starvation of auxotrophs) or by prolonged incubation with antibiotics (transcriptional and translational inhibitors) most cytoplasmic polysomes disappear and ribosomes which accumulate, upon pressure-dissociation, yield subunits displaying a normal reassociation kinetics. Such reassociated particles, however, dissociate again in the course of high-speed centrifugation unless fixed with glutaraldehyde. Polysomes from multiplying bacteria, upon incubation with RNase and French Press treatment, yield subunits reassociating into monosomes that are labile to low-speed centrifugation; this effect is still prevented by glutaraldehyde fixation. Ribosomes produced under different experimental conditions can, thus, be classified according to the 2 criteria of (a) reassociation capacity after pressure dissociation, and (b) resistance of reassociated particles to high- and low-speed centrifugation, into four distinct groups (non reassociating subunits, and subunits forming low- speed-labile, high-speed-labile and centrifugation-resistant ribosomes).
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Communicated by F. Gros
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Cocito, C. Pressure dissociation of bacterial ribosomes and reassociation of ribosomal subunits. Molec. Gen. Genet. 162, 43–50 (1978). https://doi.org/10.1007/BF00333849
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DOI: https://doi.org/10.1007/BF00333849