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Volume 169, Issue 2

Methionyl-tRNA formyltransferase utilizes 10-formyldihydrofolate as an alternative substrate and impacts antifolate drug action Open Access

Abstract

Methionyl-tRNA formyltransferase (Fmt)-mediated formylation of Met-tRNA to fMet-tRNA is crucial for efficient initiation of translation in bacteria and the eukaryotic organelles. Folate dehydrogenase-cyclohydrolase (FolD), a bifunctional enzyme, carries out conversion of 5,10-methylene tetrahydrofolate (5,10-CH-THF) to 10-formyl-THF (10-CHO-THF), a metabolite utilized by Fmt as a formyl group donor. In this study, using and approaches, we show that 10-CHO-DHF may also be utilized by Fmt as an alternative substrate (formyl group donor) to formylate Met-tRNA. Dihydrofolate (DHF) formed as a by-product in the assay was verified by LC-MS/MS analysis. FolD-deficient mutants and Fmt over-expressing strains were more sensitive to trimethoprim (TMP) than the ∆ strain, suggesting that the domino effect of TMP leads to inhibition of protein synthesis and strain growth. Antifolate treatment to showed a decrease in the reduced folate species (THF, 5,10-CH-THF, 5-CH-THF, 5,10-CH-THF and 5-CHO-THF) and increase in the oxidized folate species (folic acid and DHF). In cells, 10-CHO-DHF and 10-CHO-folic acid were enriched in the stationary phase. This suggests that 10-CHO-DHF is a bioactive metabolite in the folate pathway for generating other folate intermediates and fMet-tRNA.

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  • Accepted:
  • Published Online:
Keyword(s): 10-CHO-DHF , 10-CHO-THF , DHF , Fmt , folate pathway , one-carbon metabolic pathway and THF
Funding
This study was supported by the:
  • Tata Trusts (IISc), India
  • Department of Biotechnology, Ministry of Science and Technology, India
    • Principle Award Recipient: UmeshVarshney
  • Science and Education Research Board, New Delhi, India
    • Principle Award Recipient: UmeshVarshney
© 2023 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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2023-02-06
2024-05-10
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Methionyl-tRNA formyltransferase utilizes 10-formyldihydrofolate as an alternative substrate and impacts antifolate drug action
Microbiology 169, 001297 (2023); https://doi.org/10.1099/mic.0.001297
/content/journal/micro/10.1099/mic.0.001297
/content/journal/micro/10.1099/mic.0.001297
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