Open AccessTotal reconstitation of active large ribosomal subunlts of the tbermoaddophllk arcfaaebacteriumSulfolobus solfataricus
Author(s) -
Paola Londei,
Joaquı́n Teixidó,
Marco Acca,
Piero Cammarano,
Ricardo Amils
Publication year - 1986
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/14.5.2269
Subject(s) - sulfolobus solfataricus , biology , sulfolobus , ribosomal rna , 50s , thermophile , incubation , ribosomal protein , protein subunit , spermine , biochemistry , sulfolobus acidocaldarius , rna , microbiology and biotechnology , ribosome , archaea , enzyme , gene
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.