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Updated: Jul 21, 2026

RIBO-seq in Bacteria: a Sample Collection and Library Preparation Protocol for NGS Sequencing
Published on: August 7, 2021
Biochemical characterization of the ribosomal decoding site
1Center for Molecular Biology of RNA, Department of Molecular, Cell and Developmental Biology, Sinsheimer Laboratories, University of California, Santa Cruz, Santa Cruz, CA 95064, USA. harry@nuvolari.ucsc.edu
Abstract:
Prior to the emergence of crystal structures of the ribosome, different ribosomal functions were identified with specific regions of ribosomal RNA by biochemical and genetic approaches. In particular, three universally conserved bases of 16S rRNA, G530, A1492 and A1493, were implicated in the interaction of the incoming aminoacyl-tRNA with the 30S subunit and mRNA. The conserved region surrounding A1492 and A1493 was called the "decoding site", based on the results of chemical probing experiments and antibiotic resistance mutations. Crystallographic studies from the Ramakrishnan laboratory have now shown that G530 loop, A1492 and A1493 undergo localized conformational changes to form an RNA structure that positions these three bases to inspect the accuracy of the codon-anticodon match with high stereochemical precision, using A-minor interactions. Some results from the pre-X-ray era may provide clues to further aspects of the decoding process.
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