6Q56

Crystal structure of the B. subtilis M1A22 tRNA methyltransferase TrmK

Summary for 6Q56

• Entry DOI: 10.2210/pdb6q56/pdb
• Descriptor: tRNA (adenine(22)-N(1))-methyltransferase, NICKEL (II) ION (3 entities in total)
• Functional Keywords: methyltransferase, trna methyltransferase, rna binding protein
• Biological source: Bacillus subtilis (strain 168)
• Total number of polymer chains: 4
• Total formula weight: 104897.98

Authors

Degut, C.,Roovers, M.,Barraud, P.,Brachet, F.,Feller, A.,Larue, V.,Al Refaii, A.,Caillet, J.,Droogmans, L.,Tisne, C. (deposition date: 2018-12-07, release date: 2019-03-27, Last modification date: 2024-01-24)

Primary citation

Degut, C.,Roovers, M.,Barraud, P.,Brachet, F.,Feller, A.,Larue, V.,Al Refaii, A.,Caillet, J.,Droogmans, L.,Tisne, C.
Structural characterization of B. subtilis m1A22 tRNA methyltransferase TrmK: insights into tRNA recognition.
Nucleic Acids Res., 47:4736-4750, 2019

PubMed Abstract: 1-Methyladenosine (m1A) is a modified nucleoside found at positions 9, 14, 22 and 58 of tRNAs, which arises from the transfer of a methyl group onto the N1-atom of adenosine. The yqfN gene of Bacillus subtilis encodes the methyltransferase TrmK (BsTrmK) responsible for the formation of m1A22 in tRNA. Here, we show that BsTrmK displays a broad substrate specificity, and methylates seven out of eight tRNA isoacceptor families of B. subtilis bearing an A22. In addition to a non-Watson-Crick base-pair between the target A22 and a purine at position 13, the formation of m1A22 by BsTrmK requires a full-length tRNA with intact tRNA elbow and anticodon stem. We solved the crystal structure of BsTrmK showing an N-terminal catalytic domain harbouring the typical Rossmann-like fold of Class-I methyltransferases and a C-terminal coiled-coil domain. We used NMR chemical shift mapping to drive the docking of BstRNASer to BsTrmK in complex with its methyl-donor cofactor S-adenosyl-L-methionine (SAM). In this model, validated by methyltransferase activity assays on BsTrmK mutants, both domains of BsTrmK participate in tRNA binding. BsTrmK recognises tRNA with very few structural changes in both partner, the non-Watson-Crick R13-A22 base-pair positioning the A22 N1-atom close to the SAM methyl group.

PubMed: 30931478
DOI: 10.1093/nar/gkz230

Experimental method

X-RAY DIFFRACTION (2 Å)