Partially modified tRNAs for the study of tRNA maturation and function

Schultz, Sarah K.; Kothe, Ute

doi:10.1016/bs.mie.2021.06.007

Cited by 5 publications

(2 citation statements)

References 71 publications

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“…As ThiI has few sequence constraints for modifying U8-containing tRNAs and instead recognizes the overall L-shaped structure ( 41 ), this strategy can be used to introduce s 4 U8 within a wide selection of tRNAs. Thus, the here described preparation of fluorescently labeled, partially modified tRNA for kinetics studies lays the foundation for study of additional tRNA modification enzymes or other tRNA-interacting enzymes, demonstrating another use for in vitro -modified tRNAs ( 42 ).…”

Section: Discussionmentioning

confidence: 96%

Molecular mechanism of tRNA binding by the Escherichia coli N7 guanosine methyltransferase TrmB

Schultz¹,

Meadows²,

Kothe³

2023

Journal of Biological Chemistry

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Section: Discussionmentioning

confidence: 96%

Molecular mechanism of tRNA binding by the Escherichia coli N7 guanosine methyltransferase TrmB

Schultz¹,

Meadows²,

Kothe³

2023

Journal of Biological Chemistry

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“…As ThiI has few sequence constraints for modifying U8-containing tRNAs and instead recognizes the overall L-shaped structure (39), this strategy can be used to introduce s 4 U8 within a wide selection of tRNAs. Thus, the here described preparation of fluorescently labelled, partially-modified tRNA for kinetics studies lays the foundation for study of additional tRNA modification enzymes or other tRNA-interacting enzymes, demonstrating another use for in vitro modified tRNAs (40).…”

Section: Discussionmentioning

confidence: 96%

Molecular mechanism of tRNA binding by theEscherichia coliN7 guanosine methyltransferase TrmB

Schultz

Meadows

Kothe

2022

Preprint

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Among the large and diverse collection of tRNA modifications, 7-methylguanosine is frequently found in the tRNA variable loop at position 46. This modification is introduced by the TrmB enzyme, which is conserved in bacteria and eukaryotes. Complementing the report of various phenotypes for different organisms lacking TrmB homologs, we report here hydrogen peroxide sensitivity for theEscherichia coli ΔtrmBknockout strain. To gain insight into the molecular mechanism of tRNA binding byE. coliTrmB in real-time, we developed a new assay based on introducing a 4-thiouridine modification at position 8 ofin vitrotranscribed tRNAPheenabling us to fluorescently labelled this unmodified tRNA. Using rapid kinetic stopped-flow measurements with this fluorescent tRNA, we examined the interaction of wild-type and single substitution variants of TrmB with tRNA. Our results reveal the role of SAM for rapid and stable tRNA binding, the rate-limiting nature of m7G46 catalysis for tRNA release, and the importance of residues R26, T127 and R155 across the entire surface of TrmB for tRNA binding.

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Modified Nucleosides as RNA Components. Structure, Biological Role and Drug Design

Drenichev,

Zenchenko,

Alexeev

2023

RNA Technologies

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Partially modified tRNAs for the study of tRNA maturation and function

Cited by 5 publications

References 71 publications

Molecular mechanism of tRNA binding by the Escherichia coli N7 guanosine methyltransferase TrmB

Molecular mechanism of tRNA binding by the Escherichia coli N7 guanosine methyltransferase TrmB

Molecular mechanism of tRNA binding by theEscherichia coliN7 guanosine methyltransferase TrmB

Modified Nucleosides as RNA Components. Structure, Biological Role and Drug Design

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