Structure of a protein L23-RNA complex located at the A-site domain of the ribosomal peptidyl transferase centre.
Protein L23 from the ribosome of Escherichia coli is the primary ribosomal product cross-linked to affinity-labelled puromycin; it lies, therefore, within the A-site domain of the peptidyl transferase centre on the 50 S subunit. We have characterized this functional domain by isolating and sequencing the RNA binding site of protein L23; it consists of two main fragments of 25 and 105 nucleotides that strongly interact and are separated by 172 nucleotides in the primary sequence. The higher-order structure of the RNA moiety was probed by chemical reagents, and by single-strand and double-strand-specific ribonucleases; a secondary structural model and a tertiary structural interaction are proposed on the basis of these data that are compatible with phylogenetic sequence comparisons. Several nucleotides exhibited altered chemical reactivity, both lower and higher, in the presence of protein L23, thereby implicating a large proportion of the RNA structure in the protein binding. The sites were located mainly at the extremities of the helices and at nucleotides that were putatively bulged out from the helices. The RNA moiety and an adjacent excised fragment contain several highly conserved sequences and a modified adenosine. Such sequences constitute important functional domains of the RNA and may contribute to the putative role of this RNA region in the peptidyl transferase centre.
D000217:Acyltransferases / Q000235:genetics*; D001345:Autoradiography; D001483:Base Sequence; D001665:Binding Sites; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000032:analysis; D029968:Escherichia coli Proteins; D046911:Macromolecular Substances; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D010458:Peptidyl Transferases / Q000235:genetics*; D014176:Protein Biosynthesis; D012335:RNA, Ribosomal / Q000378:metabolism; D012260:Ribonucleases; D012269:Ribosomal Proteins / Q000378:metabolism; D012270:Ribosomes / Q000032:analysis*
D029968:Escherichia coli Proteins; D046911:Macromolecular Substances; D012335:RNA, Ribosomal; D012269:Ribosomal Proteins; C043816:ribosomal protein L23, E coli; D000217:Acyltransferases; D010458:Peptidyl Transferases; D012260:Ribonucleases