TY - JOUR
T1 - Phosphorothioate Modification of mRNA Accelerates the Rate of Translation Initiation to Provide More Efficient Protein Synthesis
AU - Kawaguchi, Daisuke
AU - Kodama, Ayumi
AU - Abe, Naoko
AU - Takebuchi, Kei
AU - Hashiya, Fumitaka
AU - Tomoike, Fumiaki
AU - Nakamoto, Kosuke
AU - Kimura, Yasuaki
AU - Shimizu, Yoshihiro
AU - Abe, Hiroshi
N1 - Funding Information:
This work was supported by the Japan Society for the Promotion of Science (JSPS) [Grant-in-Aid for Scientific Research (B), 16H04178 to H.A.; Grant-in-Aid for Early-Career Scientists, 18K14357 to N. A.] and CREST [JPMJCR18S1 to H.A.] from the Japan Science and Technology Agency (JST). We thank Assoc. Prof. Natsuhisa Oka (Gifu University, Japan) and Prof. Kiyoshi Asai (The University of Tokyo, Japan) for helpful discussions.
Funding Information:
This work was supported by the Japan Society for the Promotion of Science (JSPS) [Grant‐in‐Aid for Scientific Research (B), 16H04178 to H.A.; Grant‐in‐Aid for Early‐Career Scientists, 18K14357 to N. A.] and CREST [JPMJCR18S1 to H.A.] from the Japan Science and Technology Agency (JST). We thank Assoc. Prof. Natsuhisa Oka (Gifu University, Japan) and Prof. Kiyoshi Asai (The University of Tokyo, Japan) for helpful discussions.
Publisher Copyright:
© 2020 Wiley-VCH GmbH
PY - 2020/9/28
Y1 - 2020/9/28
N2 - 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.
AB - 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.
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U2 - 10.1002/anie.202007111
DO - 10.1002/anie.202007111
M3 - Article
C2 - 32627275
AN - SCOPUS:85089299426
VL - 59
SP - 17403
EP - 17407
JO - Angewandte Chemie - International Edition
JF - Angewandte Chemie - International Edition
SN - 1433-7851
IS - 40
ER -