Gcn4 misregulation reveals a direct role for the evolutionary conserved EKC/KEOPS in the t6A modification of tRNAs

被引：74

作者：

Daugeron, Marie-Claire ^{[1
,2
,3
]}

Lenstra, Tineke L. ^{[4
]}

Frizzarin, Martina ^{[1
,2
,5
]}

El Yacoubi, Basma ^{[6
]}

Liu, Xipeng ^{[1
]}

Baudin-Baillieu, Agnes ^{[7
]}

Lijnzaad, Philip ^{[4
]}

Decourty, Laurence ^{[8
,9
]}

Saveanu, Cosmin ^{[8
,9
]}

Jacquier, Alain ^{[8
,9
]}

Holstege, Frank C. P. ^{[4
]}

de Crecy-Lagard, Valerie ^{[6
]}

van Tilbeurgh, Herman ^{[10
]}

Libri, Domenico ^{[1
,2
]}

机构：

[1] CNRS, Ctr Genet Mol, LEA Lab Nucl RNA Metab, FRE3144, F-91190 Gif Sur Yvette, France

[2] Aarhus Univ, Ctr mRNP Biogenesis & Metab, DK-8000 Aarhus C, Denmark

[3] Univ Paris 11, F-91405 Orsay, France

[4] Univ Med Ctr Utrecht, Dept Physiol Chem, NL-3584 CG Utrecht, Netherlands

[5] Univ Padua, Dept Biol Chem, I-35100 Padua, Italy

[6] Univ Florida, Dept Microbiol & Cell Sci, Gainesville, FL 32611 USA

[7] Univ Paris 11, Inst Genet & Microbiol, UMR8621, F-91405 Orsay, France

[8] Inst Pasteur, Unite Genet Interact Macromol, F-75015 Paris, France

[9] Inst Pasteur, Unite Genet Interact Macromol, F-75015 Paris, France

[10] Univ Paris 11, Inst Biochim & Biophys Mol & Cellulaire, F-91405 Orsay, France

来源：

NUCLEIC ACIDS RESEARCH | 2011年 / 39卷 / 14期

基金：

美国国家卫生研究院;

关键词：

EUKARYOTIC TRANSLATION INITIATION; TRANSFER RIBONUCLEIC-ACID; SACCHAROMYCES-CEREVISIAE; ESCHERICHIA-COLI; IN-VITRO; TELOMERE REPLICATION; MODIFIED NUCLEOSIDE; PROTEIN-SYNTHESIS; KEOPS COMPLEX; YEAST;

D O I：

10.1093/nar/gkr178

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

070307 [化学生物学]; 071010 [生物化学与分子生物学];

摘要：

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 (t(6)A(37)) 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 t(6)A(37) 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.

引用

页码：6148 / 6160

页数：13

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