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Updated: Nov 21, 2025

Dual DNA Rulers to Study the Mechanism of Ribosome Translocation with Single-Nucleotide Resolution
Published on: July 8, 2019
High-Resolution DNA Dual-Rulers Reveal a New Intermediate State in Ribosomal Frameshifting
Yujia Mao1, Ran Lin2, Shoujun Xu1
1Department of Chemistry, University of Houston, 3585 Cullen Blvd, Houston, TX 77204, USA.
Abstract:
Ribosomal frameshifting is an important pathway used by many viruses for protein synthesis that involves mRNA translocation of various numbers of nucleotides. Resolving the mRNA positions with subnucleotide precision will provide critical mechanistic information that is difficult to obtain with current techniques. We report a method of high-resolution DNA rulers with subnucleotide precision and the discovery of new frameshifting intermediate states on mRNA containing a GA7 G motif. Two intermediate states were observed with the aid of fusidic acid, one at the "0" reading frame and the other near the "-1" reading frame, in contrast to the "-2" and "-1" frameshifting products found in the absence of the antibiotic. We termed the new near-"-1" intermediate the Post(-1*) state because it was shifted by approximately half a nucleotide compared to the normal "-1" reading frame at the 5'-end. This indicates a ribosome conformation that is different from the conventional model of three reading frames. Our work reveals uniquely precise mRNA motions and subtle conformational changes that will complement structural and fluorescence studies.
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