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  Structural model of the M7G46 methyltransferase TrmB in complex with tRNA

Blersch, K. F., Burchert, J.-P., August, S.-C., Welp, L., Neumann, P., Köster, S., et al. (2021). Structural model of the M7G46 methyltransferase TrmB in complex with tRNA. RNA Biology, In Press. doi:10.1080/15476286.2021.1925477.

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Blersch, K. F., Author
Burchert, J.-P., Author
August, S.-C., Author
Welp, L.1, Author              
Neumann, P., Author
Köster, S., Author
Urlaub, H.2, Author              
Ficner, R., Author
Affiliations:
1Research Group of Bioanalytical Mass Spectrometry, MPI for Biophysical Chemistry, Max Planck Society, ou_578613              
2Research Group of Bioanalytical Mass Spectrometry, MPI for biophysical chemistry, Max Planck Society, ou_578613              

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Free keywords: Trna modification; 7-methylguanosine; m7G; trmb; trm8
 Abstract: TrmB belongs to the class I S-adenosylmethionine (SAM)-dependent methyltransferases (MTases) and introduces a methyl group to guanine at position 7 (m7G) in tRNA. In tRNAs m7G is most frequently found at position 46 in the variable loop and forms a tertiary base pair with C13 and U22, introducing a positive charge at G46. The TrmB/Trm8 enzyme family is structurally diverse, as TrmB proteins exist in a monomeric, homodimeric, and heterodimeric form. So far, the exact enzymatic mechanism, as well as the tRNA-TrmB crystal structure is not known. Here we present the first crystal structures of B. subtilis TrmB in complex with SAM and SAH. The crystal structures of TrmB apo and in complex with SAM and SAH have been determined by X-ray crystallography to 1.9 Å (apo), 2.5 Å (SAM), and 3.1 Å (SAH). The obtained crystal structures revealed Tyr193 to be important during SAM binding and MTase activity. Applying fluorescence polarization, the dissociation constant Kd of TrmB and tRNAPhe was determined to be 0.12 µM ± 0.002 µM. Luminescence-based methyltransferase activity assays revealed cooperative effects during TrmB catalysis with half-of-the-site reactivity at physiological SAM concentrations. Structural data retrieved from small-angle x-ray scattering (SAXS), mass-spectrometry of cross-linked complexes, and molecular docking experiments led to the determination of the TrmB-tRNAPhe complex structure.

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Language(s): eng - English
 Dates: 2021-05-192021
 Publication Status: Published in print
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 Rev. Type: Peer
 Identifiers: DOI: 10.1080/15476286.2021.1925477
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Title: RNA Biology
Source Genre: Journal
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Pages: 14 Volume / Issue: - Sequence Number: In Press Start / End Page: - Identifier: -