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Science 10 April 1998:
Vol. 280. no. 5361, pp. 286 - 289
DOI: 10.1126/science.280.5361.286
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Reports

Ribosome-Catalyzed Peptide-Bond Formation with an A-Site Substrate Covalently Linked to 23S Ribosomal RNA

Rachel Green, * Christopher Switzer, Harry F. Noller dagger

In the ribosome, the aminoacyl-transfer RNA (tRNA) analog 4-thio-dT-p-C-p-puromycin crosslinks photochemically with G2553 of 23S ribosomal RNA (rRNA). This covalently linked substrate reacts with a peptidyl-tRNA analog to form a peptide bond in a peptidyl transferase-catalyzed reaction. This result places the conserved 2555 loop of 23S rRNA at the peptidyl transferase A site and suggests that peptide bond formation can occur uncoupled from movement of the A-site tRNA. Crosslink formation depends on occupancy of the P site by a tRNA carrying an intact CCA acceptor end, indicating that peptidyl-tRNA, directly or indirectly, helps to create the peptidyl transferase A site.

R. Green and H. F. Noller, Center for Molecular Biology of RNA, Sinsheimer Laboratories, University of California, Santa Cruz, CA 95064, USA.
C. Switzer, Department of Chemistry, University of California, Riverside, CA 92521, USA.
*   Present address: Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

dagger    To whom correspondence should be addressed. E-mail: harry{at}nuvolari.ucsc.edu

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