Abstract
Photoreactive 3-(4’-benzoylphenyl)propionyl-Phe-tRNA was bound specifically to the P-site or the A-site of the E. coli ribosome. Photoreaction led to high yield crosslinking between the aminoacyl-terminus of Phe-tRNA and 23S RNA. The site of reaction was identified by hybridization and by taking advantage of the fact that reverse transcriptase stops one base before a modified nucleotide. The nucleotides labeled at the P-site were identified as A-2451 and C-2452. A-site specific labeling occurred at U-2584 and U-2585. Although distant in the primary sequence, these residues lie in close proximity within the central loop of domain V according to the secondary structure model of 23S RNA. Only antibiotics known to act at the peptidyl transferase site strongly inhibit the photoaffinity reaction, thus providing further evidence for the specificity of the labeling. The demonstration of specific, high yield crosslinks at the level of 23S RNA strongly supports the involvement of an RNA-catalyzed activity in the peptidyl transferase reaction.
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Kuechler, E., Steiner, G., Barta, A. (1989). Dissection of the Ribosomal Peptidyl Transferase Center by Photoaffinity Labeling: P- And A-Sites Are Located At Different Positions in Domain V of 23 S RNA. In: Nielsen, P.E. (eds) Photochemical Probes in Biochemistry. NATO ASI Series, vol 272. Springer, Dordrecht. https://doi.org/10.1007/978-94-009-0925-0_10
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DOI: https://doi.org/10.1007/978-94-009-0925-0_10
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