eIF5A promotes +1 programmed ribosomal frameshifting in Euplotes octocarinatus

Yu Xiao; Jia Li; Ruanlin Wang; Yajiao Fan; Xiaxia Han; Yuejun Fu; Paula Alepuz; Wei Wang; Aihua Liang

doi:10.1016/j.ijbiomac.2023.127743

eIF5A promotes +1 programmed ribosomal frameshifting in Euplotes octocarinatus

Int J Biol Macromol. 2024 Jan;254(Pt 1):127743. doi: 10.1016/j.ijbiomac.2023.127743. Epub 2023 Oct 29.

Authors

Yu Xiao¹, Jia Li¹, Ruanlin Wang², Yajiao Fan¹, Xiaxia Han¹, Yuejun Fu¹, Paula Alepuz³, Wei Wang⁴, Aihua Liang⁵

Affiliations

¹ Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan 030006, China.
² Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan 030006, China. Electronic address: rlwang@sxu.edu.cn.
³ Instituto de Biotecnología y Biomedicina (Biotecmed) and Departamento de Bioquímica y Biología Molecular, Universitat de València, Spain.
⁴ Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan 030006, China. Electronic address: gene@sxu.edu.cn.
⁵ Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan 030006, China. Electronic address: aliang@sxu.edu.cn.

PMID: 38287569
DOI: 10.1016/j.ijbiomac.2023.127743

Abstract

Programmed ribosomal frameshifting (PRF) exists in all branches of life that regulate gene expression at the translational level. The single-celled eukaryote Euplotes exhibit high frequency of PRF. However, the molecular mechanism of modulating Euplotes PRF remains largely unknown. Here, we identified two novel eIF5A genes, eIF5A1 and eIF5A2, in Euplotes octocarinatus and found that the Eo-eIF5A2 gene requires a -1 PRF to produce complete protein product. Although both Eo-eIF5As showed significant structural similarity with yeast eIF5A, neither of them could functionally replace yeast eIF5A. Eo-eIF5A knockdown inhibited +1 PRF of the η-tubulin gene. Using an in vitro reconstituted translation system, we found that hypusinated Eo-eIF5A (Eo-eIF5A^H) can promote +1 PRF at the canonical AAA_UAA frameshifting site of Euplotes. The results showed eIF5A is a novel trans-regulator of PRF in Euplotes and has an evolutionary conserved role in regulating +1 PRF in eukaryotes.

Keywords: Euplotes octocarinatus; Programmed ribosomal frameshifting; eIF5A.

MeSH terms

Euplotes* / genetics
Euplotes* / metabolism
Frameshifting, Ribosomal* / genetics
Saccharomyces cerevisiae / genetics