Coordinated translation initiation determines ‐1 programmed ribosomal frameshifting efficiency of chromosomal genes to impact on cell fitness

FEBS Journal, vol. 293, pp. 4374–4396

Abstract

The conserved -1 programmed ribosomal frameshifting (-1PRF), essential for eubacteria and RNA viruses, is thought to be exclusively regulated during translation elongation. One of the two required -1PRF cis-acting elements is a frameshifting stimulatory element (FSE), which pauses the elongating ribosome to stimulate -1PRF and is consecutively deformed. Consequently, the remaining hypothesis involves the coordination between translation initiation and FSE reformation, which facilitates the -1PRF events in trailing ribosomes. Here, we create a system that allows a tunable translation initiation rate based on the MS2 bacteriophage coat protein translation initiation hairpin to assess this conjecture. We discovered a conserved negative correlation between translation initiation rate and -1PRF efficiency in eubacteria and mammalian cells. Mechanistic exploration using Escherichia coli (E. coli) shows that a higher initiation rate reduces the frequencies of FSE reformation, resulting in lower -1PRF efficiency. The role of translation initiation-mediated -1PRF was examined in an E. coli cellular copA/copA(Z) -1PRF event. The results showed that a higher translation initiation rate leads to lower -1PRF efficiency, sensitizing E. coli to copper stress. Together, these results suggest that translation initiation is an additional mechanism to regulate -1PRF to coordinate optimal protein synthesis for cell fitness, further providing the basis to investigate the interplay between translation initiation and elongation.

Authors 11

1. Sih‐Yun Tai

   National Cheng Kung University

   Affiliation as printed

   Department of Biochemistry and Molecular Biology, College of Medicine National Cheng Kung University Tainan Taiwan

2. Masayuki Hashimoto

   National Cheng Kung University

   Affiliation as printed

   Institute of Basic Medical Sciences, College of Medicine National Cheng Kung University Tainan Taiwan

   Institute of Molecular Medicine, College of Medicine National Cheng Kung University Tainan Taiwan

3. Yi‐Fang Zhuang

   National Cheng Kung University

   Affiliation as printed

   Department of Biochemistry and Molecular Biology, College of Medicine National Cheng Kung University Tainan Taiwan

4. Hao‐Wen Liang

   National Cheng Kung University

   Affiliation as printed

   Department of Biochemistry and Molecular Biology, College of Medicine National Cheng Kung University Tainan Taiwan

5. Hsin‐Pei Huang

   National Cheng Kung University

   Affiliation as printed

   Department of Biochemistry and Molecular Biology, College of Medicine National Cheng Kung University Tainan Taiwan

   Institute of Basic Medical Sciences, College of Medicine National Cheng Kung University Tainan Taiwan

6. The‐Phuong Nguyen

   Hanoi National University of Education · VNU University of Education · National Cheng Kung University

   Affiliation as printed

   Department of Microbiology and Biotechnology, Faculty of Biology Hanoi National University of Education Hanoi Vietnam

   Institute of Molecular Medicine, College of Medicine National Cheng Kung University Tainan Taiwan

7. Meng‐Han Tu

   National Cheng Kung University

   Affiliation as printed

   Department of Biochemistry and Molecular Biology, College of Medicine National Cheng Kung University Tainan Taiwan

8. Shih-Cheng Chen

   National Health Research Institutes · National Cheng Kung University

   Affiliation as printed

   Department of Biochemistry and Molecular Biology, College of Medicine National Cheng Kung University Tainan Taiwan

   National Institute of Cancer Research National Health Research Institutes Taipei Taiwan

9. Whei-Fen Wu

   National Taiwan University

   Affiliation as printed

   Department of Agricultural Chemistry, College of Bio‐Resource and Agriculture National Taiwan University Taipei Taiwan

10. René C. L. Olsthoorn Aachen

    Leiden University

    Affiliation as printed

    Leiden Institute of Chemistry Leiden University Leiden The Netherlands

11. Chien‐Hung Yu corresponding

    National Cheng Kung University

    Affiliation as printed

    Department of Biochemistry and Molecular Biology, College of Medicine National Cheng Kung University Tainan Taiwan

    Institute of Basic Medical Sciences, College of Medicine National Cheng Kung University Tainan Taiwan