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Title: Gcn4 misregulation reveals a direct role for the evolutionary conserved $$\mathrm{EKC/KEOPS}$$ in the t6A modification of t$$\mathrm{RNA}$$s

Journal Article · · Nucleic Acids Research
DOI:https://doi.org/10.1093/nar/gkr178· OSTI ID:1904624
 [1];  [2];  [3];  [4];  [5];  [6];  [2];  [7];  [7];  [7];  [2];  [4];  [6];  [8]
  1. Centre National de la Recherche Scientifique (CNRS), Gif sur Yvette (France). LEA Laboratory of Nuclear RNA metabolism; Aarhus University (Denmark); University Paris-Sud, Orsay (France)
  2. Utrecht University (Netherlands). University Medical Center
  3. Centre National de la Recherche Scientifique (CNRS), Gif sur Yvette (France). LEA Laboratory of Nuclear RNA metabolism; Aarhus University (Denmark); University of Padova (Italy)
  4. University of Florida, Gainesville, FL (United States)
  5. Centre National de la Recherche Scientifique (CNRS), Gif sur Yvette (France). LEA Laboratory of Nuclear RNA metabolism
  6. University Paris-Sud, Orsay (France)
  7. Institute Pasteur, CNRS, Paris (France). Unite de Genetique des Interactions Macromoleculaires
  8. Centre National de la Recherche Scientifique (CNRS), Gif sur Yvette (France). LEA Laboratory of Nuclear RNA metabolism; Aarhus University (Denmark)
+ Show Author Affiliations

The EKC/KEOPS complex is universally conserved in Archaea and Eukarya and has been implicated in several cellular processes, including transcription, telomere homeostasis and genomic instability. However, the molecular function of the complex has remained elusive so far. We analyzed the transcriptome of EKC/KEOPS mutants and observed a specific profile that is highly enriched in targets of the Gcn4p transcriptional activator. GCN4 expression was found to be activated at the translational level in mutants via the defective recognition of the inhibitory upstream ORFs (uORFs) present in its leader. We show that EKC/KEOPS mutants are defective for the N6-threonylcarbamoyl adenosine modification at position 37 (t6A37) of tRNAs decoding ANN codons, which affects initiation at the inhibitory uORFs and provokes Gcn4 de-repression. Structural modeling reveals similarities between Kae1 and bacterial enzymes involved in carbamoylation reactions analogous to t6A37 formation, supporting a direct role for the EKC in tRNA modification. These findings are further supported by strong genetic interactions of EKC mutants with a translation initiation factor and with threonine biosynthesis genes. Overall, our data provide a novel twist to understanding the primary function of the EKC/KEOPS and its impact on several essential cellular functions like transcription and telomere homeostasis.

Research Organization:
Univ. of Florida, Gainesville, FL (United States)
Sponsoring Organization:
USDOE Office of Science (SC); Centre National pour la Recherche Scientifique (CNRS); Agence Nationale pour la Recherche (ANR); Netherlands Organization of Scientific Research (NWO); Netherlands Bioinformatics Centre (NBIC); National Institutes of Health (NIH)
Grant/Contract Number:
FG02-07ER64498; ANR-09-BLAN-0349-03; ANR-08-JCJC-0019-01; 021.002.035; 817.02.015; 050.71.057; 911.06.009; 016.108.607; R01 GM70641-01
OSTI ID:
1904624
Journal Information:
Nucleic Acids Research, Vol. 39, Issue 14; ISSN 0305-1048
Publisher:
Oxford University PressCopyright Statement
Country of Publication:
United States
Language:
English

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Cited By (18)

Bilaminar Structure of the Developing Stapedial Footplate in the Mouse —A Histological Study Using A Light Microscope— journal January 1987
tRNA thiolation links translation to stress responses in Saccharomyces cerevisiae journal January 2015
Molecular basis for t6A modification in human mitochondria journal February 2020
Structure of a reaction intermediate mimic in t6A biosynthesis bound in the active site of the TsaBD heterodimer from Escherichia coli journal February 2021
The ATP-mediated formation of the YgjD–YeaZ–YjeE complex is required for the biosynthesis of tRNA t6A in Escherichia coli journal January 2015
KEOPS complex promotes homologous recombination via DNA resection journal April 2019
Conformational communication mediates the reset step in t6A biosynthesis journal May 2019
Structure and Mechanism of a Bacterial t6A Biosynthesis System journal April 2018
Transcriptome-wide analysis of roles for tRNA modifications in translational regulation journal June 2017
A familial case of Galloway-Mowat syndrome due to a novel TP53RK mutation: a case report journal July 2018
Drosophilap53-related protein kinase is required for PI3K/TOR pathway-dependent growth journal March 2013
Loss of a Conserved tRNA Anticodon Modification Perturbs Cellular Signaling journal August 2013
The Human EKC/KEOPS Complex Is Recruited to Cullin2 Ubiquitin Ligases by the Human Tumour Antigen PRAME journal August 2012
A Genetic Investigation of the KEOPS Complex in Halophilic Archaea journal August 2012
The emerging role of complex modifications of tRNALysUUU in signaling pathways journal January 2015
Global translational impacts of the loss of the tRNA modification t6A in yeast journal January 2016
KAE1 Allelic Variants Affect TORC1 Activation and Fermentation Kinetics in Saccharomyces cerevisiae journal July 2019
Diversity of the biosynthesis pathway for threonylcarbamoyladenosine (t 6 A), a universal modification of tRNA journal December 2014

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