Total reconstitation of active large ribosomal subunlts of the tbermoaddophllk arcfaaebacteriumSulfolobus solfataricus
- 1 January 1986
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 14 (5) , 2269-2285
- https://doi.org/10.1093/nar/14.5.2269
Abstract
The large ribosomal subunit of the extremely thermoacidophilic archaebacterium Sulfolobus solfataricus has been reconstituted from the completely dissociated RNA and proteins by a two-step incubation procedure at high temperatures. Successful reconstitution requires a preliminary incubation of the ribosomal components for 45 min at 65 degrees C, followed by a second heat-treatment at 80 degrees C for 60 min. Structural reassembly depends upon high concentrations of K+ (300-400 mM) and Mg2+ (20-40 mM) ions. In addition, complete recovery of subunit function stringently requires the presence of a polyamine, thermine (or spermine). The reconstituted archaebacterial subunits are essentially indistinguishable from the native ones by a number of structural and functional criteria.Keywords
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