RESUMEN
Modified uridine containing taurine, 5-taurinomethyluridine (τm5U), is found at the anticodon first position of mitochondrial (mt-)transfer RNAs (tRNAs). Previously, we reported that τm5U is absent in mt-tRNAs with pathogenic mutations associated with mitochondrial diseases. However, biogenesis and physiological role of τm5U remained elusive. Here, we elucidated τm5U biogenesis by confirming that 5,10-methylene-tetrahydrofolate and taurine are metabolic substrates for τm5U formation catalyzed by MTO1 and GTPBP3. GTPBP3-knockout cells exhibited respiratory defects and reduced mitochondrial translation. Very little τm5U34 was detected in patient's cells with the GTPBP3 mutation, demonstrating that lack of τm5U results in pathological consequences. Taurine starvation resulted in downregulation of τm5U frequency in cultured cells and animal tissues (cat liver and flatfish). Strikingly, 5-carboxymethylaminomethyluridine (cmnm5U), in which the taurine moiety of τm5U is replaced with glycine, was detected in mt-tRNAs from taurine-depleted cells. These results indicate that tRNA modifications are dynamically regulated via sensing of intracellular metabolites under physiological condition.
Asunto(s)
ARN de Transferencia/metabolismo , Taurina/deficiencia , Uridina/análogos & derivados , Animales , Proteínas Portadoras/fisiología , Gatos , Preescolar , Femenino , Proteínas de Unión al GTP/genética , Proteínas de Unión al GTP/fisiología , Células HEK293 , Células HeLa , Humanos , Mitocondrias/metabolismo , Enfermedades Mitocondriales/genética , ARN de Transferencia/química , Proteínas de Unión al ARN , Uridina/biosíntesisRESUMEN
The relative growth rate of plant cells in vitro is considerably affected by initial cell density. This troublesome effect has interfered with the establishment of efficient plant cell culture systems, especially when only a small number of cells are expected to survive, such as in the genetic transformation of cells under antibiotic selection. To improve the recovery of antibiotic-resistant cells, we examined the use of the peptide plant hormone phytosulfokine (PSK), which has been shown to promote cellular growth and development in vitro. The addition of PSK to selective media increased the recovery of transformed callus from Agrobacterium-infected carrot hypocotyl explants from 7% to 39%, which is more than a fivefold improvement over the control. Most calluses developed into normal plantlets with cotyledons and primary roots and, eventually, formed foliage leaves. Thus, chemical nursing using PSK shows promise as a tool for basic research in plant biology and biotechnological applications.