Escherichia coli methionine-tRNAi/methionyl tRNA synthetase pairs induced protein initiation of interest (PII) expression

Jung Min Kim, Han Yong Lee, Jinho Jung

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

The precise regulatory role in protein synthesis by facilitating interactions with mRNA codons for various tRNA modifications is unclear. We previously reported that enhanced green fluorescent protein (GFP) reduced enhanced GFP mRNA expression in human methionine-conjugated initiator tRNA (tRNAi)/tRNA synthetase pairs under methionine-deficient conditions. Here, we investigated the effect of non-formylated methionine-conjugated Escherichia coli tRNAi on the synthesis of the protein initiation of interest (PII) in HeLa cells under intracellular L-methionine levels. We found that E. coli methionine-tRNAi counteracts human methionine-tRNAi, indicating that E. coli methionyl tRNA synthetase can induce enhanced GFP expression due to increased stability of enhanced GFP mRNA. Both complexes could support translation initiation without being employed to introduce methionine residues in the subsequent elongation steps. The results indicated that E. coli methionine-tRNAi could offset human methionine-tRNAi, and E. coli methionine-tRNAi/methionyl tRNA synthetase pairs can drive enhanced GFP mRNA expression. Unlike the human methionine-tRNAi/methionyl tRNA synthetase pairs that were used as a positive control, the non-formylated E. coli methionine-tRNAi/methionyl tRNA synthetase pairs reduced the expression of enhanced GFP mRNA, resulting in reduced HeLa cell survival. Using tRNAs functions causes of heterologous origin, such as from prokaryotes, and modified, to enhance or suppress the synthesis of specific proteins in eukaryotic organisms into the potential may possess a more prominent advantage of E. coli methionine-tRNAi as approaches that can control PII. This study provides new insights on the E. coli methionine- tRNAi/methionyl tRNA synthetase pair induced PII synthesis and the relative viability of cells could pave the way to regulate ecological/biological systems.

Original languageEnglish
Article number80
JournalApplied Biological Chemistry
Volume65
Issue number1
DOIs
Publication statusPublished - 2022 Dec

Bibliographical note

Funding Information:
This research was supported by Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education(2022R1A6A3A01086819)(JMK). This work was also supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No.2022-0662) (HYL).

Funding Information:
We thank Dr. Baik Lin Seong (Yonsei University) for valuable advice. We also thank Dr. Sang-Hyeon Kang (iNtRON Biotechnology) for E. coli tRNA technical supports. The expression vector encoding the human initiator tRNA gene was a gift from Dr. Sung Hun Kim (Yonsei University). We are also grateful to Dr. Joo Hee Chung of the KBSI (Seoul) for HPLC and MALDI TOF analysis.

Publisher Copyright:
© 2022, The Author(s).

Keywords

  • E. coli initiator tRNA
  • E. coli methionyl tRNA synthetase
  • Formylated methionine
  • Non-formylated methionine
  • Transfer RNA
  • Translation initiation

ASJC Scopus subject areas

  • General Biochemistry,Genetics and Molecular Biology
  • Organic Chemistry

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