The conserved Wobble uridine tRNA thiolase Ctu1–Ctu2 is required to maintain genome integrity

Dewez, Monique; Bauer, Fanélie; Dieu, Marc; Raes, Martine; Vandenhaute, Jean; Hermand, Damien

doi:10.1073/pnas.0709404105

Cited by 131 publications

(161 citation statements)

References 51 publications

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“…The role of thiomodification of tRNAs remains unclear. Thiolation of tRNAs may stabilize codon-anticodon interactions on the ribosome to improve translational efficiency (17,32,33). However, whether the Urm1 pathway contributes to efficient translation of endogenous proteins has not been determined.…”

Section: Discussionmentioning

confidence: 99%

“…Next, MOCS3 adenylates the C terminus of Urm1, followed by the transfer of sulfur to the terminal glycine of Urm1 (15). Urm1 associates with the thiouridylases ATPBD3 (also known as CTU1 in Homo sapiens and Ncs6p in S. cerevisiae) and CTU2 (Ncs2p in S. cerevisiae), which mediate the thiolation of wobble uridines in tRNA Lys(UUU) , tRNA Gln(UUG) , and tRNA Glu(UUC) (9)(10)(11)(12)(13)(16)(17)(18).…”

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confidence: 99%

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Role of the ubiquitin-like protein Urm1 as a noncanonical lysine-directed protein modifier

Veen

Schorpp

Schlieker

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

171

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The ubiquitin (Ub)-related modifier Urm1 functions as a sulfur carrier in tRNA thiolation by means of a mechanism that requires the formation of a thiocarboxylate at the C-terminal glycine residue of Urm1. However, whether Urm1 plays an additional role as a Ublike protein modifier remains unclear. Here, we show that Urm1 is conjugated to lysine residues of target proteins and that oxidative stress enhances protein urmylation in both Saccharomyces cerevisiae and mammalian cells. Similar to ubiquitylation, urmylation involves a thioester intermediate and results in the formation of a covalent peptide bond between Urm1 and its substrates. In contrast to modification by canonical Ub-like modifiers, however, conjugation of Urm1 involves a C-terminal thiocarboxylate of the modifier. We have confirmed that the peroxiredoxin Ahp1 is such a substrate in S. cerevisiae and found that Urm1 targets a specific lysine residue of Ahp1 in vivo. In addition, we have identified several unique substrates in mammalian cells and show that Urm1 targets at least two pathways on oxidant treatment. First, Urm1 is appended to lysine residues of three components that function in its own pathway (i.e., MOCS3, ATPBD3, and CTU2). Second, Urm1 is conjugated to the nucleocytoplasmic shuttling factor cellular apoptosis susceptibility protein. Thus, Urm1 has a conserved dual role by integrating the functions of prokaryotic sulfur carriers with those of eukaryotic protein modifiers of the Ub family.posttranslational modification | hydrogen peroxide | nuclear transport | Uba4 | USP15

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Role of the ubiquitin-like protein Urm1 as a noncanonical lysine-directed protein modifier

Veen

Schorpp

Schlieker

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

171

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“…In yeast, inactivation of wobble uridine modification enzymes can impair translational decoding, as measured by decreased translation of relevant 'codon-runs' 26,27 . To investigate the possible effects of the extensive wobble U mis-modification observed in the Alkbh8 − / − mice, we generated reporter plasmids containing both Renilla (Rluc; internal reference) and firefly (Fluc) luciferase genes, in which Fluc, but not Rluc, translation depends on in-frame translation with a codon-run of ten identical codons (Fig.…”

Section: Evolutionary Distribution Of Alkbh8 and Mchm 5 U Modificationsmentioning

confidence: 99%

ALKBH8-mediated formation of a novel diastereomeric pair of wobble nucleosides in mammalian tRNA

et al. 2011

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Mammals have nine different homologues (ALKBH1-9) of the Escherichia coli DNA repair demethylase AlkB. ALKBH2 is a genuine DNA repair enzyme, but the in vivo function of the other ALKBH proteins has remained elusive. It was recently shown that ALKBH8 contains an additional transfer RNA (tRNA) methyltransferase domain, which generates the wobble nucleoside 5-methoxycarbonylmethyluridine (mcm 5 U) from its precursor 5-carboxymethyluridine (cm 5 U). In this study, we report that (R)-and (S)-5-methoxycarbonylhydroxymethyluridine (mchm 5 U), hydroxylated forms of mcm 5 U, are present in mammalian tRNA UCG Arg , and tRNA UCC Gly , respectively, representing the first example of a diastereomeric pair of modified RNA nucleosides. Through in vitro and in vivo studies, we show that both diastereomers of mchm 5 U are generated from mcm 5 U, and that the AlkB domain of ALKBH8 specifically hydroxylates mcm 5 U into (S)-mchm 5 U in tRNA UCC Gly . These findings expand the function of the ALKBH oxygenases beyond nucleic acid repair and increase the current knowledge on mammalian wobble uridine modifications and their biogenesis.

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“…In fact, Rep2 is one of the proteins that is supposed to be not well translated in some Elongator mutants. 20 However, we do not observe any decrease in the amount of Rep2 that is present in cells of Elongator or tRNA methylation mutants (Fig. 3D).…”

Section: Discussionmentioning

confidence: 78%

“…To confirm that the effect measured with the reporter strain was on MBF-dependent transcription, we isolated RNA from wild type, Dyox1 and Drep2 cells as well as from Diki3/Delp1 and Dctu1 cells. We used Dctu1 cells since the Ctu1-Ctu2 complex catalyzes the thiolation at carbon 2 of U 34 (s 2 U 34 ) 20 and the modification of specific tRNAs is sequentially made by Trm112 complex and the Ctu1-Ctu2 complex. In fact, although Dctu1 is not included in Table 1 containing the lowest YFP/ FSC strains from our screening, Dctu1 strain had also a low YFP/FSC ratio (0.66).…”

Section: Trna Methylation and Elongator Mutants Have Impaired Mbf-depmentioning

confidence: 99%

A functional genome-wide genetic screening identifies new pathways controlling the G1/S transcriptional wave

et al. 2016

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The Schizosaccharomyces pombe MBF complex activates the transcription of genes required for DNA synthesis and S phase. The MBF complex contains several proteins, including the core components Cdc10, Res1 and Res2, the co-repressor proteins Yox1 and Nrm1 and the co-activator Rep2. It has recently been shown how MBF is regulated when either the DNA damage or the DNA synthesis checkpoints are activated. However, how MBF is regulated in a normal unperturbed cell cycle is still not well understood. We have set up a genome-wide genomic screen searching for global regulators of MBF. We have crossed our knock-out collection library with a reporter strain that allows the measurement of MBF activity in live cells by flow cytometry. We confirm previously known regulators of MBF and show that COP9/ signalosome and tRNA methyltransferases also regulate MBF activity.

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The conserved Wobble uridine tRNA thiolase Ctu1–Ctu2 is required to maintain genome integrity

Cited by 131 publications

References 51 publications

Role of the ubiquitin-like protein Urm1 as a noncanonical lysine-directed protein modifier

Role of the ubiquitin-like protein Urm1 as a noncanonical lysine-directed protein modifier

ALKBH8-mediated formation of a novel diastereomeric pair of wobble nucleosides in mammalian tRNA

A functional genome-wide genetic screening identifies new pathways controlling the G1/S transcriptional wave

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