title: From bacterial to human dihydrouridine synthase: automated structure determination.
abstract: The reduction of uridine to dihydrouridine at specific positions in tRNA is catalysed by dihydrouridine synthase (Dus) enzymes. Increased expression of human dihydrouridine synthase 2 (hDus2) has been linked to pulmonary carcinogenesis, while its knockdown decreased cancer cell line viability, suggesting that it may serve as a valuable target for therapeutic intervention. Here, the X-ray crystal structure of a construct of hDus2 encompassing the catalytic and tRNA-recognition domains (residues 1-340) determined at 1.9 Å resolution is presented. It is shown that the structure can be determined automatically by phenix.mr_rosetta starting from a bacterial Dus enzyme with only 18% sequence identity and a significantly divergent structure. The overall fold of the human Dus2 is similar to that of bacterial enzymes, but has a larger recognition domain and a unique three-stranded antiparallel β-sheet insertion into the catalytic domain that packs next to the recognition domain, contributing to domain-domain interactions. The structure may inform the development of novel therapeutic approaches in the fight against lung cancer.
authors.affiliation: Department of Biology, The University of York, Heslington, York YO10 5DD, England.
authors.forename: Fiona
authors.initials: F
authors.lastname: Whelan
authors.affiliation: York Structural Biology Laboratory, Department of Chemistry, The University of York, Heslington, York YO10 5DD, England.
authors.forename: Huw T
authors.initials: HT
authors.lastname: Jenkins
authors.affiliation: Division of Structural Biology, University of Oxford, Headington, Oxford OX3 7BN, England.
authors.forename: Samuel C
authors.initials: SC
authors.lastname: Griffiths
authors.affiliation: Gene Center and Department of Biochemistry, Ludwig-Maximilians-University Munich, Feodor-Lynen-Strasse 25, 81377 Munich, Germany.
authors.forename: Robert T
authors.initials: RT
authors.lastname: Byrne
authors.affiliation: York Structural Biology Laboratory, Department of Chemistry, The University of York, Heslington, York YO10 5DD, England.
authors.forename: Eleanor J
authors.initials: EJ
authors.lastname: Dodson
authors.affiliation: York Structural Biology Laboratory, Department of Chemistry, The University of York, Heslington, York YO10 5DD, England.
authors.forename: Alfred A
authors.initials: AA
authors.lastname: Antson
mesh_terms: D001419:Bacteria / Q000737:chemistry / Q000201:enzymology / Q000378:metabolism; D001665:Binding Sites; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D006801:Humans; D008175:Lung Neoplasms / Q000201:enzymology; D008958:Models, Molecular; D010088:Oxidoreductases / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D012343:RNA, Transfer / Q000378:metabolism
chemical_list: D012343:RNA, Transfer; D010088:Oxidoreductases; C526987:tRNA dihydrouridine synthase-2, human
keywords: MR-Rosetta; X-ray crystallography; dihydrouridine synthase; lung cancer; tRNA modification