title: Structural and functional insights into tRNA binding and adenosine N1-methylation by an archaeal Trm10 homologue.
abstract: Purine nucleosides on position 9 of eukaryal and archaeal tRNAs are frequently modified in vivo by the post-transcriptional addition of a methyl group on their N1 atom. The methyltransferase Trm10 is responsible for this modification in both these domains of life. While certain Trm10 orthologues specifically methylate either guanosine or adenosine at position 9 of tRNA, others have a dual specificity. Until now structural information about this enzyme family was only available for the catalytic SPOUT domain of Trm10 proteins that show specificity toward guanosine. Here, we present the first crystal structure of a full length Trm10 orthologue specific for adenosine, revealing next to the catalytic SPOUT domain also N- and C-terminal domains. This structure hence provides crucial insights in the tRNA binding mechanism of this unique monomeric family of SPOUT methyltransferases. Moreover, structural comparison of this adenosine-specific Trm10 orthologue with guanosine-specific Trm10 orthologues suggests that the N1 methylation of adenosine relies on additional catalytic residues.
authors.affiliation: Structural Biology Brussels, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussel, Belgium Structural Biology Research Center, VIB, Pleinlaan 2, 1050 Brussel, Belgium.
authors.forename: Bart
authors.initials: B
authors.lastname: Van Laer
authors.affiliation: Institut de Recherches Microbiologiques Jean-Marie Wiame, Avenue E. Gryson 1, 1070 Bruxelles, Belgium.
authors.forename: Martine
authors.initials: M
authors.lastname: Roovers
authors.affiliation: Structural Biology Brussels, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussel, Belgium Structural Biology Research Center, VIB, Pleinlaan 2, 1050 Brussel, Belgium Department of Cell Biochemistry, University of Groningen, Nijenborgh 7, Groningen 9747 AG, Netherlands.
authors.forename: Lina
authors.initials: L
authors.lastname: Wauters
authors.affiliation: International Institute of Molecular and Cell Biology in Warsaw, Trojdena 4 St, 02-109 Warsaw, Poland Institute of Molecular Biology and Biotechnology, Faculty of Biology, Adam Mickiewicz University, 61-614 Poznan, Poland.
authors.forename: Joanna M
authors.initials: JM
authors.lastname: Kasprzak
authors.affiliation: International Institute of Molecular and Cell Biology in Warsaw, Trojdena 4 St, 02-109 Warsaw, Poland.
authors.forename: Michal
authors.initials: M
authors.lastname: Dyzma
authors.affiliation: Structural Biology Brussels, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussel, Belgium Structural Biology Research Center, VIB, Pleinlaan 2, 1050 Brussel, Belgium.
authors.forename: Egon
authors.initials: E
authors.lastname: Deyaert
authors.affiliation: Structural Biology Brussels, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussel, Belgium Structural Biology Research Center, VIB, Pleinlaan 2, 1050 Brussel, Belgium.
authors.forename: Ranjan
authors.initials: R
authors.lastname: Kumar Singh
authors.affiliation: Laboratoire de Microbiologie, Université libre de Bruxelles, 12 Rue des Professeurs Jeener et Brachet, 6041 Gosselies, Belgium.
authors.forename: André
authors.initials: A
authors.lastname: Feller
authors.affiliation: International Institute of Molecular and Cell Biology in Warsaw, Trojdena 4 St, 02-109 Warsaw, Poland Institute of Molecular Biology and Biotechnology, Faculty of Biology, Adam Mickiewicz University, 61-614 Poznan, Poland.
authors.forename: Janusz M
authors.identifier: http://orcid.org/0000-0002-6633-165X
authors.initials: JM
authors.lastname: Bujnicki
authors.affiliation: Laboratoire de Microbiologie, Université libre de Bruxelles, 12 Rue des Professeurs Jeener et Brachet, 6041 Gosselies, Belgium ldroogma@ulb.ac.be.
authors.forename: Louis
authors.initials: L
authors.lastname: Droogmans
authors.affiliation: Structural Biology Brussels, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussel, Belgium Structural Biology Research Center, VIB, Pleinlaan 2, 1050 Brussel, Belgium Wim.Versees@vib-vub.be.
authors.forename: Wim
authors.initials: W
authors.lastname: Versées
mesh_terms: D000241:Adenosine / Q000737:chemistry / Q000378:metabolism*; D019843:Archaeal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D008745:Methylation; D008958:Models, Molecular; D062105:Molecular Docking Simulation; D017434:Protein Structure, Tertiary; D012343:RNA, Transfer / Q000737:chemistry / Q000378:metabolism*; D012358:RNA, Transfer, Met / Q000737:chemistry / Q000378:metabolism; D053838:Scattering, Small Angle; D016945:Sulfolobus acidocaldarius / Q000201:enzymology*; D014961:X-Ray Diffraction; D012359:tRNA Methyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D019843:Archaeal Proteins; D012358:RNA, Transfer, Met; D012343:RNA, Transfer; D012359:tRNA Methyltransferases; D000241:Adenosine