Structure of the EMAPII domain of human aminoacyl-tRNA synthetase complex reveals evolutionary dimer mimicry

Renault, Louis; Kerjan, Pierre; Pasqualato, Sebastiano; Menetrey, J.; Robinson, Jean Charles; Kawaguchi, Shin‐ichi; Vassylyev, Dmitry G.; Yokoyama, Shigeyuki; Mirande, Marc; Cherfils, Jacqueline

doi:10.1093/emboj/20.3.570

Cited by 64 publications

(79 citation statements)

References 52 publications

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“…Crystal structures reveal that the C-terminal or EMAPII (endothelial monocyte-activating polypeptide) domain of p43 forms a truncated dimeric oligonucleotide-binding fold. This serves as a nonspecific RNA binding domain (29,32), which may function in trans to recruit tRNAs for specific enzyme(s) within the multisynthetase complex. This would result in enhancement of catalytic efficiencies (7,33).…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, formation of the p43 dimer is believed to occur by interaction of their N termini (8). One can project a size for the connected monomers based on the dimensions of the crystal structure of the C-terminal half of p43 (29). The 5 nm distance between spots 2 and 3 on the three-dimensional structure of the multisynthetase complex is within the range that could be spanned by the p43 dimer.…”

Section: Isolation Of Multisynthetase Complex Via a Novel Methods And mentioning

confidence: 99%

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A Three-dimensional Working Model of the Multienzyme Complex of Aminoacyl-tRNA Synthetases Based on Electron Microscopic Placements of tRNA and Proteins

Wolfe

Warrington

Treadwell

et al. 2005

Journal of Biological Chemistry

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It has become evident that the process of protein synthesis is performed by many cellular polypeptides acting in concert within the structural confines of protein complexes. In multicellular eukaryotes, one of these assemblies is a multienzyme complex composed of eight proteins that have aminoacyl-tRNA synthetase activities as well as three non-synthetase proteins (p43, p38, and p18) with diverse functions. This study uses electron microscopy and three-dimensional reconstruction to explore the arrangement of proteins and tRNA substrates within this "core" multisynthetase complex. Binding of unfractionated tRNA establishes that these molecules are widely distributed on the exterior of the structure. Binding of gold-labeled tRNALeu places leucyl-tRNA synthetase and the bifunctional glutamyl-/prolyl-tRNA synthetase at the base of this asymmetric "V"-shaped particle. A stable cell line has been produced that incorporates hexahistidine-labeled p43 into the multisynthetase complex. Using a gold-labeled nickel-nitrilotriacetic acid probe, the polypeptides of the p43 dimer have been located along one face of the particle. The results of this and previous studies are combined into an initial three-dimensional working model of the multisynthetase complex. This is the first conceptualization of how the protein constituents and tRNA substrates are arrayed within the structural confines of this multiprotein assembly.It has been known for many years that a "core" complex of aminoacyltRNA synthetases can be isolated from all multicellular eukaryotes (1). Its characteristic composition is three non-synthetase proteins and eight polypeptides having nine aminoacyl-tRNA synthetase activities. With the enzymes abbreviated as the three-letter amino acid name plus RS for the enzyme, the components are ArgRS, AspRS, GluProRS, GlnRS, IleRS, LeuRS, LysRS, MetRS, p43, p38, and p18. Several of these are known dimers, so the total polypeptide count in the multisynthetase complex is at least fifteen. This is a particularly intriguing protein assembly as all of the enzymes catalyze the same reaction, albeit with individual and highly specific substrates. The reaction catalyzed is the covalent attachment of an amino acid to either the 2Ј-or 3Ј-hydroxyl of the 3Ј-terminal adenosine of tRNA. Recent increased interest in these enzymes has been generated by studies indicating that they have roles in a variety of other cellular processes (2-4). Such activities range from cytokine-mediated chemoattraction to priming of reverse transcription of the human immunodeficiency virus, type 1 genome. These enzymes are also viewed as targets in the development of new antimicrobial agents (5). The multisynthetase complex also contains three non-synthetase proteins. The first of these, p18, has been shown to interact with EF-1␥, which may facilitate transfer of aminoacyl-tRNAs to the ribosome (6). Data from yeast two-hybrid screens (7), in vitro binding assays (8), and deletion analysis (9) indicate that p38 functions as a scaffolding protein and appears to be...

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Section: Discussionmentioning

confidence: 99%

Section: Isolation Of Multisynthetase Complex Via a Novel Methods And mentioning

confidence: 99%

A Three-dimensional Working Model of the Multienzyme Complex of Aminoacyl-tRNA Synthetases Based on Electron Microscopic Placements of tRNA and Proteins

Wolfe

Warrington

Treadwell

et al. 2005

Journal of Biological Chemistry

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“…To understand the multifunctionality and regulation of the components of the multi-ARS complex, it is important to determine the three-dimensional structures of the components as well as the whole complex. So far, only the three-dimensional structures of the partial domains of the complex-forming ARSs such as glutamylprolyl-tRNA synthetase (10,11), aspartyltRNA synthetase (12), and AIMP1/p43 (13,14) have been elucidated. Here, we report the crystal structure of full-length AIMP3 and the residues and location required for the interaction with ATM.…”

mentioning

confidence: 99%

Determination of Three-dimensional Structure and Residues of the Novel Tumor Suppressor AIMP3/p18 Required for the Interaction with ATM

Kim

Park

Choi

et al. 2008

Journal of Biological Chemistry

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Although AIMP3/p18 is normally associated with the multitRNA synthetase complex via its specific interaction with methionyl-tRNA synthetase, it also works as a tumor suppressor by interacting with ATM, the upstream kinase of p53. To understand the molecular interactions of AIMP3 and the mechanisms involved, we determined the crystal structure of AIMP3 at 2.0-Å resolution and identified its potential sites of interaction with ATM. AIMP3 contains two distinct domains linked by a 7-amino acid (Lys 57 -Ser 63 ) peptide, which contains a 3 10 helix. The 56-amino acid N-terminal domain consists of two helices into which three antiparallel ␤ strands are inserted, and the 111-amino acid C-terminal domain contains a bundle of five helices (Thr 64 -Tyr 152 ) followed by a coiled region (Pro 153 -Leu 169 ). Structural analyses revealed homologous proteins such as yeast glutamyl-tRNA synthetase, Arc1p, EF1B␥, and glutathione S-transferase and suggested two potential molecular binding sites. Moreover, mutations at the C-terminal putative binding site abolished the interaction between AIMP3 and ATM and the ability of AIMP3 to activate p53. Thus, this work identified the two potential molecular interaction sites of AIMP3 and determined the residues critical for its tumor-suppressive activity through the interaction with ATM.Although aminoacyl-tRNA synthetases (ARSs) 4 catalyze the ligation of specific amino acids to their cognate tRNAs, they are multifunctional proteins that are involved in the regulation of diverse signal pathways (1). Several ARSs of higher eukaryotes form an intriguing macromolecular protein complex with three non-enzymatic factors, designated AIMPs (ARS-interacting multifunctional proteins), which are also multiply involved in various biological processes. Of these factors, AIMP1/p43 is secreted as a cytokine that functions in immune response (2, 3), angiogenesis (4), and wound-healing processes (5) and also as a hormone that regulates glucose metabolism (6). AIMP2/p38 has been shown to mediate transforming growth factor-␤ signaling for c-Myc down-regulation (7). AIMP3/p18 binds to the multi-ARS complex by specifically interacting with methionyltRNA synthetase (MRS) and is translocated to the nucleus to activate p53 through the interaction with kinases such as ATM (ataxia telangiectasia mutated) and ATR (ATM-and Rad3-related) in response to DNA damage (8) or oncogenic stress (9). To understand the multifunctionality and regulation of the components of the multi-ARS complex, it is important to determine the three-dimensional structures of the components as well as the whole complex. So far, only the three-dimensional structures of the partial domains of the complex-forming ARSs such as glutamylprolyl-tRNA synthetase (10, 11), aspartyltRNA synthetase (12), and AIMP1/p43 (13, 14) have been elucidated. Here, we report the crystal structure of full-length AIMP3 and the residues and location required for the interaction with ATM. EXPERIMENTAL PROCEDURESCloning, Expression, and Purification of Human AIMP3-The cDN...

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“…The p43 protein is located in the middle of the complex (9) and is associated with arginyl-tRNA synthetase via its N-terminal region (10). Its C-terminal domain contains an OB-fold, which is responsible for the interaction with tRNA (11,12) and facilitates the catalytic activity of the bound enzyme (10). Interestingly, p43 is also secreted to work as a proinflammatory cytokine (13,14).…”

mentioning

confidence: 99%

p38 is essential for the assembly and stability of macromolecular tRNA synthetase complex: Implications for its physiological significance

Kim

Kang

Lee

et al. 2002

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

110

113

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Mammalian tRNA synthetases form a macromolecular complex with three nonenzyme factors: p43, p38, and p18. Here we introduced a mutation within the mouse p38 gene to understand its functional significance for the formation of the multi-tRNA synthetase complex. The complex was completely disintegrated by the deficiency of p38. In addition, the protein levels and catalytic activities of the component enzymes and cofactors were severely decreased. A partial truncation of the p38 polypeptide separated the associated components into different subdomains. The mutant mice showed lethality within 2 days of birth. Thus, this work provides the first evidence, to our knowledge, that p38 is essential for the structural integrity of the multi-tRNA synthetase complex and mouse viability.aminoacyl-tRNA synthetase ͉ macromolecular protein complex ͉ gene trap ͉ protein-protein interaction A minoacyl-tRNA synthetases (ARSs) are essential enzymes catalyzing the ligation of their cognate amino acids and tRNAs. Eight different enzymes of higher eukaryotes form a macromolecular complex with three nonsynthetase factors: p43, p38, and p18 (1-3). Although this complex was first reported more than two decades ago, the functional reason for their molecular assembly and the structural organization of the components still remain unknown.Recently, much information has been obtained about the associations and interactions of the component proteins. The assembly of the complex is mediated by heat-shock protein 90 (4) and involves protein-protein interactions via the unique noncatalytic peptides attached to each of the component enzymes (5-7) and their catalytic core domains (8). It was expected that the three associating factors would also contribute to the complex formation. Among the three auxiliary factors, the interaction and function of p43 have been best elucidated. The p43 protein is located in the middle of the complex (9) and is associated with arginyl-tRNA synthetase via its N-terminal region (10). Its C-terminal domain contains an OB-fold, which is responsible for the interaction with tRNA (11, 12) and facilitates the catalytic activity of the bound enzyme (10). Interestingly, p43 is also secreted to work as a proinflammatory cytokine (13,14). The functions of the two other factors are less understood. p38 interacts with many components of the complex (2, 15). To understand the in vivo functional significance of p38 in the formation of the multi-ARS complex, we mutated the p38 structural gene in the mouse and investigated its effects on the cellular stability of the multi-ARS complex and the component proteins. Deletion analyses of p38 were also conducted to map the organization of the component proteins within the complex.

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Structure of the EMAPII domain of human aminoacyl-tRNA synthetase complex reveals evolutionary dimer mimicry

Cited by 64 publications

References 52 publications

A Three-dimensional Working Model of the Multienzyme Complex of Aminoacyl-tRNA Synthetases Based on Electron Microscopic Placements of tRNA and Proteins

A Three-dimensional Working Model of the Multienzyme Complex of Aminoacyl-tRNA Synthetases Based on Electron Microscopic Placements of tRNA and Proteins

Determination of Three-dimensional Structure and Residues of the Novel Tumor Suppressor AIMP3/p18 Required for the Interaction with ATM

p38 is essential for the assembly and stability of macromolecular tRNA synthetase complex: Implications for its physiological significance

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