title: A common tRNA modification at an unusual location: the discovery of wyosine biosynthesis in mitochondria.
abstract: Establishment of the early genetic code likely required strategies to ensure translational accuracy and inevitably involved tRNA post-transcriptional modifications. One such modification, wybutosine/wyosine is crucial for translational fidelity in Archaea and Eukarya; yet it does not occur in Bacteria and has never been described in mitochondria. Here, we present genetic, molecular and mass spectromery data demonstrating the first example of wyosine in mitochondria, a situation thus far unique to kinetoplastids. We also show that these modifications are important for mitochondrial function, underscoring their biological significance. This work focuses on TyW1, the enzyme required for the most critical step of wyosine biosynthesis. Based on molecular phylogeny, we suggest that the kinetoplastids pathways evolved via gene duplication and acquisition of an FMN-binding domain now prevalent in TyW1 of most eukaryotes. These findings are discussed in the context of the extensive U-insertion RNA editing in trypanosome mitochondria, which may have provided selective pressure for maintenance of mitochondrial wyosine in this lineage.
authors.affiliation: Department of Microbiology and The Center for RNA Biology, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Sample
authors.affiliation: Institute of Parasitology, Biology Centre and Faculty of Sciences, University of South Bohemia, 37005 České Budějovice (Budweis), Czech Republic.
authors.forename: Luděk
authors.initials: L
authors.lastname: Kořený
authors.affiliation: Department of Microbiology and The Center for RNA Biology, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Zdeněk
authors.initials: Z
authors.lastname: Paris
authors.affiliation: Rieveschl Laboratories for Mass Spectrometry, Department of Chemistry, University of Cincinnati, Cincinnati, OH 45221, USA.
authors.forename: Kirk W
authors.initials: KW
authors.lastname: Gaston
authors.affiliation: Department of Microbiology and The Center for RNA Biology, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Mary Anne T
authors.initials: MA
authors.lastname: Rubio
authors.affiliation: Department of Microbiology and The Center for RNA Biology, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Ian M C
authors.initials: IM
authors.lastname: Fleming
authors.affiliation: Department of Microbiology and The Center for RNA Biology, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Scott
authors.initials: S
authors.lastname: Hinger
authors.affiliation: Institute of Parasitology, Biology Centre and Faculty of Sciences, University of South Bohemia, 37005 České Budějovice (Budweis), Czech Republic.
authors.forename: Eva
authors.initials: E
authors.lastname: Horáková
authors.affiliation: Rieveschl Laboratories for Mass Spectrometry, Department of Chemistry, University of Cincinnati, Cincinnati, OH 45221, USA.
authors.forename: Patrick A
authors.initials: PA
authors.lastname: Limbach
authors.affiliation: Institute of Parasitology, Biology Centre and Faculty of Sciences, University of South Bohemia, 37005 České Budějovice (Budweis), Czech Republic Canadian Institute For Advanced Research, Toronto, ON M5G 1Z8, Canada.
authors.forename: Julius
authors.initials: J
authors.lastname: Lukeš
authors.affiliation: Department of Microbiology and The Center for RNA Biology, The Ohio State University, Columbus, OH 43210, USA Rieveschl Laboratories for Mass Spectrometry, Department of Chemistry, University of Cincinnati, Cincinnati, OH 45221, USA Ohio State Biochemistry Program, The Ohio State University, Columbus, Ohio 43210, USA alfonzo.1@osu.edu.
authors.forename: Juan D
authors.initials: JD
authors.lastname: Alfonzo
mesh_terms: D006151:Guanosine / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D008928:Mitochondria / Q000201:enzymology*; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D012343:RNA, Transfer / Q000737:chemistry / Q000378:metabolism*; D014346:Trypanosoma brucei brucei / Q000201:enzymology* / Q000235:genetics
chemical_list: D015800:Protozoan Proteins; D006151:Guanosine; C012100:wyosine; D012343:RNA, Transfer
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