Cloning and characterization of cDNA encoding a human arginyl-tRNA synthetase

Girjes, Adeeb A.; Hobson, Karen; Chen, Philip; Lavin, Martin F.

doi:10.1016/0378-1119(95)00502-w

Cited by 15 publications

(20 citation statements)

References 13 publications

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“…RNA analysis and cDNA cloning support the contention that only one form of ArgRS mRNA exists in human cells (19,30). N-Terminal amino acid sequence analysis of the purified free ArgRS from rat liver suggests that lowmolecular weight ArgRS is probably a distinct translational form, rather than a proteolytically derived product (12).…”

Section: Discussionmentioning

confidence: 88%

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Two Forms of Human Cytoplasmic Arginyl-tRNA Synthetase Produced from Two Translation Initiations by a Single mRNA

et al. 2006

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Human cytoplasmic arginyl-tRNA synthetase (ArgRS) is a component of a macromolecular complex consisting of at least nine tRNA synthetases and three auxiliary proteins. In mammalian cells, ArgRS is present as a free protein as well as a component of the complex. Via an alignment of ArgRSs from different vertebrates, the genes encoding full-length human cytoplasmic ArgRS and an N-terminal 72-amino acid deletion mutant (hcArgRS and ∆NhcArgRS, respectively) were subcloned and expressed in Escherichia coli. The two ArgRS products were expressed as a soluble protein in E. coli. The level of production of ∆NhcArgRS in E. coli and its specific activity were higher than those for hcArgRS. By Western blot analysis, using an antibody against the purified ∆NhcArgRS, the two forms of ArgRS were detected in three human cell types. The 5′-end cDNA sequence, as confirmed by 5′RACE (5′-rapid amplification of cDNA ends), contained three start codons. Through mutation of the three codons, the two human cytoplasmic ArgRSs were found to be produced in different amounts, indicating that they resulted from two different translation initiation events. Here we show evidence that two forms of human cytoplasmic ArgRS were produced from two translational initiations by a single mRNA.

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Section: Discussionmentioning

confidence: 88%

“…Only one form of ArgRS cDNA has been cloned from CHO, mouse, 5HSY-5Y, 293T, and HepG2 cells, the hcArgRS:∆NhcArgRS ratios were 2.9, 3.08, and 2.1, respectively. and human cells (18,19,30). Also, a single human cytoplasmic ArgRS mRNA of 2.2 kb has been detected by Northern blot analysis (19).…”

Section: Discussionmentioning

confidence: 99%

Two Forms of Human Cytoplasmic Arginyl-tRNA Synthetase Produced from Two Translation Initiations by a Single mRNA

et al. 2006

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“…Although the activity of RRS was enhanced about 2.5-fold by pro-EMAPII under our experimental conditions, its effect may be more significant in vivo because RRS present in the multi-protein complex would have limited accessibility to tRNA Mammalian RRS exists in two forms differing by the Nterminal extension (15). The larger RRS containing the Nterminal extension is found in the multi-synthetase complex, whereas the smaller RRS exists in a free form (18,19). The complex-associated larger RRS showed a 7-fold higher K m for the tRNA substrate than the complex-free RRS, whereas other kinetic properties were similar (15).…”

Section: Discussionmentioning

confidence: 99%

“…In the present work, we focused on the interaction between pro-EMAPII and RRS. The N-terminal 72-amino acid peptide region is only found in human (18) and hamster RRS proteins (19). We conducted deletion analysis to determine the peptide regions of pro-EMAPII and RRS responsible for the interaction.…”

Section: Screening Of Proteins Interacting With Human Pro-ema-pii-mentioning

confidence: 99%

Precursor of Pro-apoptotic Cytokine Modulates Aminoacylation Activity of tRNA Synthetase

Park¹,

Jung²,

Lee³

et al. 1999

Journal of Biological Chemistry

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Endothelial monocyte activating polypeptide II (EMA-PII) is a cytokine that is specifically induced by apoptosis.Its precursor (pro-EMAPII) has been suggested to be identical to p43, which is associated with the multi-tRNA synthetase complex. Herein, we have demonstrated that the N-terminal domain of pro-EMAPII interacts with the Nterminal extension of human cytoplasmic arginyl-tRNA synthetase (RRS) using genetic and immunoprecipitation analyses. Aminoacylation activity of RRS was enhanced about 2.5-fold by the interaction with pro-EMAPII but not with its N-or C-terminal domains alone. The N-terminal extension of RRS was not required for enzyme activity but did mediate activity stimulation by pro-EMAPII. Pro-EMAPII reduced the apparent K m of RRS to tRNA, whereas the k cat value remained unchanged. Therefore, the precursor of EMAPII is a multi-functional protein that assists aminoacylation in normal cells and releases the functional cytokine upon apoptosis.Aminoacyl-tRNA synthetases (ARSs) 1 catalyze ligation of their cognate amino acids to specific tRNAs. Although basic architecture of the core domain is well conserved among ARSs, unique peptide extensions have been found in the N-or Cterminal ends of metazoan enzymes (1-3). Although these extensions have been thought to be involved in heterologous molecular interactions, their functional significance is not yet understood.A macromolecular protein complex consisting of at least nine different ARSs has been found in higher eukaryotes (1-3). This multi-ARS complex also contains three nonsynthetase components, p18, p38, and p43 whose functions are not clear (4 -7). Among these nonsynthetase components, p43 has been proposed to be a precursor of a tumor-specific cytokine, endothelial monocyte-activating polypeptide II (EMAPII) based on over 80% sequence identity between the two proteins (6). EMAPII was originally identified in the culture medium of murine fibrosarcoma cells induced by methylcholanthrene A (8). It triggers an acute inflammatory response (9, 10) and is involved in development-related apoptosis (11).The precursor for EMAPII (pro-EMAPII) is processed at the Asp residue of ASTD/S sequence to release the C-terminal cytokine domain of 23 kDa (11). Its C-terminal domain shares homology with the C-terminal parts of methionyl-tRNA synthetases of prokaryotes, archaea and nematode, and also a yeast protein, Arc1p/G4p, which interacts with methionyl-and glutamyl-tRNA synthetases. The N-terminal domain of pro-EMAPII does not show homology to any known proteins, and its function has not been understood.EMAPII is expressed in a wide range of cell lines and normal tissues (12) and its mRNA level is unchanged during apoptosis (11) although its production is induced by apoptosis. The present work was designed to address whether pro-EMAPII is identical to p43 and to understand its function in the normal cell. The results showed that pro-EMAPII is associated with the N-terminal extension of human arginyl-tRNA synthetase (RRS), facilitating aminoacylation reaction. EXPE...

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“…To resolve this issue, the endogenous low molecular weight ArgRS was separated and purified from rat liver tissue, and an N-terminal sequence analysis suggested that it is probably a distinct translation product (11). A Northern blot analysis revealed that there is only one single transcript of approximate 2.2 kb in human cells, corresponding to the high molecular weight form of ArgRS (13). Furthermore, 5Ј RACE (Rapid Amplication of cDNA Ends) was conducted to map the 5Ј end of hcArgRS mRNA, which confirmed that there is only one mRNA encoding ArgRS in human 293T cells (14).…”

mentioning

confidence: 99%

The mRNA of Human Cytoplasmic Arginyl-tRNA Synthetase Recruits Prokaryotic Ribosomes Independently

Fang

Ruan

et al. 2014

Journal of Biological Chemistry

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Background: Two isoforms of human cytoplasmic arginyl-tRNA synthetase (hcArgRS) are produced from a single mRNA. Results: Two isoforms were found when hcArgRS was overexpressed in E. coli. Conclusion: The mRNA encoding the N-terminal extension of hcArgRS can recruit E. coli ribosomes independently. Significance: This study may provide a mechanism of alternative translational initiation of hcArgRS mRNA in human cells.

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Cloning and characterization of cDNA encoding a human arginyl-tRNA synthetase

Cited by 15 publications

References 13 publications

Two Forms of Human Cytoplasmic Arginyl-tRNA Synthetase Produced from Two Translation Initiations by a Single mRNA

Two Forms of Human Cytoplasmic Arginyl-tRNA Synthetase Produced from Two Translation Initiations by a Single mRNA

Precursor of Pro-apoptotic Cytokine Modulates Aminoacylation Activity of tRNA Synthetase

The mRNA of Human Cytoplasmic Arginyl-tRNA Synthetase Recruits Prokaryotic Ribosomes Independently

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