Identification of factors that promote biogenesis of tRNA_CGA^Ser

Abstract

A wide variety of factors are required for the conversion of pre-tRNA molecules into the mature tRNAs that function in translation. To identify factors influencing tRNA biogenesis, we previously performed a screen for strains carrying mutations that induce lethality when combined with a sup61-T47:2C allele, encoding a mutant form of . Analyzes of two complementation groups led to the identification of Tan1 as a protein involved in formation of the modified nucleoside N4-acetylcytidine (ac4C) in tRNA and Bud13 as a factor controlling the levels of ac4C by promoting TAN1 pre-mRNA splicing. Here, we describe the remaining complementation groups and show that they include strains with mutations in genes for known tRNA biogenesis factors that modify (DUS2, MOD5 and TRM1), transport (LOS1), or aminoacylate (SES1) . Other strains carried mutations in genes for factors involved in rRNA/mRNA synthesis (RPA49, RRN3 and MOT1) or magnesium uptake (ALR1). We show that mutations in not only DUS2, LOS1 and SES1 but also in RPA49, RRN3 and MOT1 cause a reduction in the levels of the altered . These results indicate that Rpa49, Rrn3 and Mot1 directly or indirectly influence biogenesis.

SSD1 suppresses phenotypes induced by the lack of Elongator-dependent tRNA modifications

“…To analyze the nucleoside composition of total tRNA, the tRNA was isolated from exponentially growing cultures at OD 600 ≈0.8 [21]. The tRNA was digested to nucleosides using nuclease P1 (Sigma-Aldrich, N8630) and bacterial alkaline phosphatase (Sigma-Aldrich, P4252) and the hydrolysate analyzed by HPLC [71, 72]. The compositions of the elution buffers were as described [72] with the difference that methanol concentration in buffer A was changed to 5% (v/v).…”

AtBUD13 affects pre‐mRNA splicing and is essential for embryo development in Arabidopsis

“…In budding yeast, only 5% of protein‐coding genes contain introns (Spingola et al ., ). Consistent with this, only a few genes with abnormal mRNA splicing, including TAN1 , MER1 and MATa1 , have been identified in the yeast bud13 ⊿ mutant cells (Scherrer and Spingola, ; Tuo et al ., ; Zhou et al ., ; Xu et al ., ). By contrast, most protein‐coding genes in higher eukaryotes harbor introns.…”

“…In the absence of Bud13p, the splicing efficiency of reporter constructs is substantially decreased both in vivo and in vitro (Gottschalk et al ., ; Dziembowski et al ., ). So far, the splicing of four pre‐mRNAs, Actin , TAN1 , MER1 and MATa1 , has been demonstrated to be controlled by Bud13p (Clark et al ., ; Khanna et al ., ; Zhou et al ., ; Wysoczanski and Zweckstetter, ; Xu et al ., ). Additionally, Bud13p has been shown to function in the formation of spliceosome and activation of the B complex (Deckert et al ., ; Bessonov et al ., ; Bao et al ., ).…”

TRMT1L-catalyzed m22G27 on tyrosine tRNA is required for efficient mRNA translation and cell survival under oxidative stress

“…The presence of dihydrouridine influences D-loop structure by promoting the formation of a hairpin with a stable stem and flexible loop and facilitates tertiary interactions that contribute to three-dimensional tRNA folding (123). In budding yeast, strains with mutations in Dus2, which modifies U20, have been shown to have reduced levels of a mutant allele of tRNA Ser-CGA, suggesting a role for D modification in imparting tRNA stability (124). Recent studies investigating the role of human DUS proteins have shown that although D is abundant across many tRNAs, only a subset of tRNA species show impaired expression levels upon DUS enzyme depletion (89).…”