Biologically active fragment of a human tRNA synthetase inhibits fluid shear stress-activated responses of endothelial cells

Tzima, Ellie; Reader, John S.; Irani-Tehrani, Mohamad; Ewalt, Karla L.; Schwartz, Martin A.; Schimmel, Paul

doi:10.1073/pnas.2436330100

Cited by 56 publications

(51 citation statements)

References 58 publications

(56 reference statements)

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“…Further studies revealed that T2-TrpRS was effective at inhibiting EC responses stimulated by either growth factors or mechanical stress (13), indicating that T2-TrpRS may interrupt these signaling systems by acting on a point common to both mechanisms. Shear stress responses inhibited by T2-TrpRS included activation of intracellular signaling pathways involving ERK and Akt, gene expression, and cell alignment (13).…”

Section: Binding Of T2-trprs To Endothelial Cells-previous Studies Idmentioning

confidence: 99%

“…Shear stress responses inhibited by T2-TrpRS included activation of intracellular signaling pathways involving ERK and Akt, gene expression, and cell alignment (13). These fundamental activities, as well as those stimulated by growth factors, are needed during angiogenesis, and therefore, inhibition of EC responses to either VEGF or shear stress could explain how T2-TrpRS inhibits angiogenesis in vivo.…”

Section: Binding Of T2-trprs To Endothelial Cells-previous Studies Idmentioning

confidence: 99%

“…Interestingly, BAECs that had been pretreated with T2-TrpRS and were subjected to long periods of shear stress failed to align their stress fibers in the direction of flow (13). Eventually cells detached from the substratum and their cell-cell contacts were visibly altered, compared with cells not treated with T2-TrpRS (13).…”

Section: Binding Of T2-trprs To Endothelial Cells-previous Studies Idmentioning

confidence: 99%

“…More recently, in vitro studies on endothelial cells (ECs) have shown that the inhibitory activity of mini-TrpRS and T2-TrpRS abrogates cellular responses involved in angiogenesis, such as motility and migration of ECs (4,6) and activation of extracellular signal-regulated kinase (ERK1/2) and Akt (13). T2-TrpRS inhibited several additional EC responses to the mechanical force of shear stress including endothelial nitric-oxide synthase activation, shear stress-induced gene expression, and cell alignment (13). While the protein binds specifically to the ends of newly formed vessels, the connection of this binding to the inhibition of angiogenic signal transduction pathways remains unknown.…”

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confidence: 99%

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VE-cadherin Links tRNA Synthetase Cytokine to Anti-angiogenic Function

Tzima

Reader

Irani-Tehrani

et al. 2005

Journal of Biological Chemistry

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A natural fragment of an enzyme that catalyzes the first step of protein synthesis-human tryptophanyltRNA synthetase (T2-TrpRS) has potent anti-angiogenic activity. A cellular receptor through which T2-TrpRS exerts its anti-angiogenic activity has not previously been identified. Here T2-TrpRS was shown to bind at intercellular junctions of endothelial cells (ECs). Using genetic knock-outs, binding was established to depend on VE-cadherin, a calcium-dependent adhesion molecule, which is selectively expressed in ECs, concentrated at adherens junctions, and is essential for normal vascular development. In contrast, T2-TrpRS binding to EC junctions was not dependent on platelet endothelial cell adhesion molecule type-1, another adhesion molecule found at EC junctions. Pull-down assays confirmed direct complex formation between T2-TrpRS and VEcadherin. Binding of T2-TrpRS inhibited VEGF-induced ERK activation and EC migration. Thus, a VE-cadherindependent pathway is proposed to link T2-TrpRS to inhibition of new blood vessel formation.During their long evolution, aminoacyl-tRNA synthetases, enzymes that catalyze the first step of protein synthesisacquired additional functions, including regulation of transcription and translation, RNA splicing, and cytokine activities in inflammatory and angiogenic signaling pathways (1, 2). Recently, fragments of the closely related human tyrosyl-tRNA and tryptophanyl-tRNA synthetase (TrpRS) 1 were discovered to regulate angiogenesis (2-6). In mammalian cells, TrpRS is activated for anti-angiogenic signaling by proteolysis or alternative splicing to give two natural isoforms-mini-TrpRS and T2-TrpRS (6). Expression of mini-TrpRS is strongly induced (along with other angiostatic factors such as IP-10 and MIG) by the anti-proliferative cytokine interferon-␥ (7-9). Mini-TrpRS and T2-TrpRS inhibit development of new vessels without affecting pre-established vasculature (4, 6, 10). Regions within mini-TrpRS that contribute to angiostatic activity have been proposed (11,12). The anti-angiogenic activity of T2-TrpRS and mini-TrpRS was demonstrated in several cell-based assays in vitro and also in vivo in the chick embryo and in the neonatal and adult mouse (4, 6). More recently, in vitro studies on endothelial cells (ECs) have shown that the inhibitory activity of mini-TrpRS and T2-TrpRS abrogates cellular responses involved in angiogenesis, such as motility and migration of ECs (4, 6) and activation of extracellular signal-regulated kinase (ERK1/2) and Akt (13). T2-TrpRS inhibited several additional EC responses to the mechanical force of shear stress including endothelial nitric-oxide synthase activation, shear stress-induced gene expression, and cell alignment (13). While the protein binds specifically to the ends of newly formed vessels, the connection of this binding to the inhibition of angiogenic signal transduction pathways remains unknown. To further elucidate the mechanism of T2-TrpRS, we sought to identify the EC target responsible for the potent anti-angiogenic activity. EXPER...

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Section: Binding Of T2-trprs To Endothelial Cells-previous Studies Idmentioning

confidence: 99%

Section: Binding Of T2-trprs To Endothelial Cells-previous Studies Idmentioning

confidence: 99%

Section: Binding Of T2-trprs To Endothelial Cells-previous Studies Idmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

VE-cadherin Links tRNA Synthetase Cytokine to Anti-angiogenic Function

Tzima

Reader

Irani-Tehrani

et al. 2005

Journal of Biological Chemistry

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“…The most critical molecular recognition events in the translation of the genetic code involve the error-free attachment of 21 amino acids to tRNAs that bear the appropriate anticodon triplets (1)(2)(3). These reactions are catalyzed by aminoacyltRNA synthetases (AARSs), 1 which are divided into two classes (I and II) based on their structures and reactivities (4).…”

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confidence: 99%

Structures of Tryptophanyl-tRNA Synthetase II from Deinococcus radiodurans Bound to ATP and Tryptophan

Buddha

Crane

2005

Journal of Biological Chemistry

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An auxiliary tryptophanyl tRNA synthetase (drTrpRS II) that interacts with nitric-oxide synthase in the radiation-resistant bacterium Deinococcus radiodurans charges tRNA with tryptophan and 4-nitrotryptophan, a specific nitration product of nitric-oxide synthase. The most critical molecular recognition events in the translation of the genetic code involve the error-free attachment of 21 amino acids to tRNAs that bear the appropriate anticodon triplets (1-3). These reactions are catalyzed by aminoacyltRNA synthetases (AARSs), 1 which are divided into two classes (I and II) based on their structures and reactivities (4). Class I synthetases attach amino acids to the 2Ј-hydroxyl group of the 3Ј-terminal adenosine of tRNA and comprise a catalytic Rossmann fold (RF) domain and a helical tRNA anticodon binding domain (TAB) (5). Two signature sequence motifs (KMSKS and (T/H)IGN) that participate in ATP binding typify class Ic synthetases (TrpRS, TyrRS, and PheRS). Some class I synthetases also incorporate an insertion domain (CP1) and participate in additional cellular functions such as RNA splicing, tRNA processing (6) RNA trafficking, and apoptosis (7).Although most AARSs are usually expressed from single copy genes to avoid the proliferation of synthetase mutations that could corrupt the genetic code, two copies of some synthetases are found in select bacteria and archea. In particular, Deinococcus radiodurans contains two tryptophanyl-tRNA synthetases (TrpRSs). Whereas D. radiodurans TrpRS (drTrpRS) I has 40% sequence identity to most single copy bacterial TrpRSs, drTrpRS II is more divergent, with only 28% sequence identity (8). We have recently shown that drTrpRS II interacts with D. radiodurans nitric oxide synthase (8, 9) and can charge tRNA with Trp and 4-nitrotryptophan, a specific nitration product of D. radiodurans nitric oxide synthase. drTrpRS II is induced during responses to DNA damage. It contains an unusual 30-residue extension, uncharacteristic of any other known prokaryotic TrpRSs (8) but homologous to the N terminus of the stationary phase stress-response protein, SurE (10).Extensive crystallographic work by Carter and colleagues (11-13) has established TrpRS from Bacillus stearothermophilus (bsTrpRS) as a paradigm for understanding the structural basis of enzymatic activity for class Ic AARSs. Crystal structures of bsTrpRS in three distinct conformations (open, closed, and closed pretransition state (pre-TS)) provide snapshots of important states in the catalytic cycle (12)(13)(14). An open conformation binds either ATP or Trp in a kinetic scheme described as "random bi-bi" (15, 16). Favorable binding interactions provided by the second substrate overcome the stability of the open state and result in domain closure. This high energy closed conformation (inferred in the closely related TyrRS from kinetic studies (17, 18)) engages the conserved KMSKS loop of the TAB domain with the ATP pyrophosphate group and brings the Trp carboxylate in close proximity of the ATP ␣-phosphate * This work was suppo...

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Temporal Genome Expression Profile Analysis During T-cell-Mediated Colitis: Identification of Novel Targets and Pathways

Fang

Zhang

Glawe

et al. 2012

Inflammatory Bowel Diseases

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T cells critically regulate IBD with T cell dependent experimental colitis models gaining favor in identifying potential pathogenic mechanisms; yet, limited understanding of specific pathogenic molecules or pathways still exists. In this study, we sought to identify changes in whole genome expression profiles using the CD4CD45Rbhi T cell transfer colitis model compared to genome expression differences from Crohn's disease tissue specimens. Colon tissue was used for histopathological and genome expression profiling analysis at 0, 2, 4, or 6 weeks after adoptive T cell transfer. We identified 1,775 genes that were significantly altered during disease progression with 361 being progressively down regulated and 341 progressively up regulated. Gene expression changes were validated by qRT-PCR confirming genome expression analysis data. Differentially expressed genes were clearly related to inflammation/immune responses but also strongly associated with metabolic, chemokine signaling, Jak-STAT signaling, and angiogenesis pathways. Ingenuity network analysis revealed 25 unique network associations that were associated with functions such as antigen presentation, cell morphology, cell-to-cell signaling and interaction, as well as nervous system development and function. Moreover, many of these genes and pathways were similarly identified in Crohn's disease specimens. These findings reveal novel, complex, and dynamic changes in gene expression that may provide useful targets for future therapeutic approaches.

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Biologically active fragment of a human tRNA synthetase inhibits fluid shear stress-activated responses of endothelial cells

Cited by 56 publications

References 58 publications

VE-cadherin Links tRNA Synthetase Cytokine to Anti-angiogenic Function

VE-cadherin Links tRNA Synthetase Cytokine to Anti-angiogenic Function

Structures of Tryptophanyl-tRNA Synthetase II from Deinococcus radiodurans Bound to ATP and Tryptophan

Temporal Genome Expression Profile Analysis During T-cell-Mediated Colitis: Identification of Novel Targets and Pathways

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