ausblenden:
Schlagwörter:
Bacterial Proteins/an [Analysis]; Base Sequence; *Cysteine Endopeptidases/an [Analysis]; Cysteine Endopeptidases/ip [Isolation & Purification]; Electrophoresis, Polyacrylamide Gel; Erythrocytes/en [Enzymology]; GroEL Protein/an [Analysis]; Hela Cells; Human; Molecular Sequence Data; *Multienzyme Complexes/an [Analysis]; Multienzyme Complexes/ip [Isolation & Purification]; *RNA/an [Analysis]; RNA/ip [Isolation & Purification]; Thermoplasma/en [Enzymology]
Zusammenfassung:
The RNA isolated from RNase-treated proteasome preparations from human erythrocytes, HeLa cells, the archaeon Thermoplasma acidophilum and also from recombinant proteasomes of T. acidophilum expressed in Escherichia coli was characterized. The RNA associated with structurally similar protein particles, namely with the two molecular chaperones, groEL from E. coli and with the thermosome from T. acidophilum, served as controls. Electrophoretic analysis on polyacrylamide gels of the radioactively end-labelled RNA revealed a very similar size distribution pattern, irrespectively of the protein particles from which they had been isolated. The predominant RNA species were in the size ranges 80 nucleotides and 120 nucleotides, respectively. Partial sequencing of their terminal regions by mobility-shift analysis revealed that, of the proteasomes from human erythrocytes, the approximately 80-nucleotide-long RNA consists of a heterogenous population of mostly tRNA species because they carried the tRNA-specific 3'-terminal sequence motif 5'-CCA-3'. The RNA in the size range 120 nucleotides isolated from the proteasomes of human erythrocytes and of T. acidophilum was also heterogeneous and displayed, in the terminal regions, a remarkable sequence similarity to the corresponding regions of the 5S rRNA from the same and different organisms. The total content of RNA of all the protein particles was quantified and found to be consistently sub-stoichiometric. All these findings strongly suggest that RNA associated with the proteasomes and with the molecular chaperones originate from the abundant cellular pool of the tRNAs and 5S rRNAs which bind non-specifically to these large protein particles.
