High‐Resolution DNA Dual‐Rulers Reveal a New Intermediate State in Ribosomal Frameshifting. (25th February 2021)

• Record Type: Journal Article
• Authors: Mao, Yujia; Lin, Ran; Xu, Shoujun; Wang, Yuhong
• Is Part Of: Chembiochem. Volume 22:Number 10(2021)
• Page Start: 1775
• Page End: 1778
• Publication Date: 2021-02-25
• Subjects: antibiotics -- atomic magnetometer -- DNA rulers -- force spectroscopy -- ribosomal frameshift; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.202000863
• Languages: English
• ISSNs: 1439-4227

Abstract

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. Abstract : Branch line : Precise DNA rulers were developed to investigate the mechanism of ribosomal frameshifting. Based on super‐resolution force spectroscopy, they revealed nucleic acid motion with subnucleotide resolution. New intermediate states were identified, one of which differed from the conventional state by less than a single nucleotide. Our work contributes towards understanding branching among different reading frames during protein synthesis.