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Phosphorothioate Modification of mRNA Accelerates the Rate of Translation Initiation to Provide More Efficient Protein Synthesis
Author(s) -
Kawaguchi Daisuke,
Kodama Ayumi,
Abe Naoko,
Takebuchi Kei,
Hashiya Fumitaka,
Tomoike Fumiaki,
Nakamoto Kosuke,
Kimura Yasuaki,
Shimizu Yoshihiro,
Abe Hiroshi
Publication year - 2020
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.202007111
Subject(s) - messenger rna , protein biosynthesis , translation (biology) , ribosome , chemistry , eukaryotic translation , microbiology and biotechnology , rna , biochemistry , biology , gene
Messenger RNAs (mRNAs) with phosphorothioate modification (PS‐mRNA) to the phosphate site of A, G, C, and U with all 16 possible combinations were prepared, and the translation reaction was evaluated using an E. coli cell‐free translation system. Protein synthesis from PS‐mRNA increased in 12 of 15 patterns when compared with that of unmodified mRNA. The protein yield increased 22‐fold when the phosphorothioate modification at A/C sites was introduced into the region from the 5′‐end to the initiation codon. Single‐turnover analysis of PS‐mRNA translation showed that phosphorothioate modification increases the number of translating ribosomes, thus suggesting that the rate of translation initiation (rate of ribosome complex formation) is positively affected by the modification. The method provides a new strategy for improving translation by using non‐natural mRNA.