title: Queuosine deficiency in eukaryotes compromises tyrosine production through increased tetrahydrobiopterin oxidation.
abstract: Queuosine is a modified pyrrolopyrimidine nucleoside found in the anticodon loop of transfer RNA acceptors for the amino acids tyrosine, asparagine, aspartic acid, and histidine. Because it is exclusively synthesized by bacteria, higher eukaryotes must salvage queuosine or its nucleobase queuine from food and the gut microflora. Previously, animals made deficient in queuine died within 18 days of withdrawing tyrosine, a nonessential amino acid, from the diet (Marks, T., and Farkas, W. R. (1997) Biochem. Biophys. Res. Commun. 230, 233-237). Here, we show that human HepG2 cells deficient in queuine and mice made deficient in queuosine-modified transfer RNA, by disruption of the tRNA guanine transglycosylase enzyme, are compromised in their ability to produce tyrosine from phenylalanine. This has similarities to the disease phenylketonuria, which arises from mutation in the enzyme phenylalanine hydroxylase or from a decrease in the supply of its cofactor tetrahydrobiopterin (BH4). Immunoblot and kinetic analysis of liver from tRNA guanine transglycosylase-deficient animals indicates normal expression and activity of phenylalanine hydroxylase. By contrast, BH4 levels are significantly decreased in the plasma, and both plasma and urine show a clear elevation in dihydrobiopterin, an oxidation product of BH4, despite normal activity of the salvage enzyme dihydrofolate reductase. Our data suggest that queuosine modification limits BH4 oxidation in vivo and thereby potentially impacts on numerous physiological processes in eukaryotes.
authors.affiliation: School of Biochemistry and Immunology, Trinity College Dublin, Dublin, Ireland.
authors.forename: Tatsiana
authors.initials: T
authors.lastname: Rakovich
authors.forename: Coilin
authors.initials: C
authors.lastname: Boland
authors.forename: Ilana
authors.initials: I
authors.lastname: Bernstein
authors.forename: Vimbai M
authors.initials: VM
authors.lastname: Chikwana
authors.forename: Dirk
authors.initials: D
authors.lastname: Iwata-Reuyl
authors.forename: Vincent P
authors.initials: VP
authors.lastname: Kelly
mesh_terms: D000818:Animals; D056945:Hep G2 Cells; D006801:Humans; D051379:Mice; D009704:Nucleoside Q / Q000235:genetics* / Q000378:metabolism*; D010084:Oxidation-Reduction; D010430:Pentosyltransferases / Q000235:genetics / Q000378:metabolism; D010649:Phenylalanine / Q000235:genetics / Q000378:metabolism; D010651:Phenylalanine Hydroxylase / Q000235:genetics / Q000378:metabolism; D010661:Phenylketonurias / Q000235:genetics / Q000378:metabolism; D011622:Pterins / Q000378:metabolism*; D012343:RNA, Transfer / Q000235:genetics / Q000378:metabolism; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D014443:Tyrosine / Q000096:biosynthesis* / Q000235:genetics*
chemical_list: D011622:Pterins; C011552:tetrahydropterin; D014443:Tyrosine; D010649:Phenylalanine; D009704:Nucleoside Q; D012343:RNA, Transfer; D010651:Phenylalanine Hydroxylase; D013762:Tetrahydrofolate Dehydrogenase; D010430:Pentosyltransferases; C020837:queuine tRNA-ribosyltransferase