PBL 2013

BIOCHEMISTRY

The molecular structure of a typical AARS and its catalytic and editing domains. The functional motifs have been highlighted and labeled appropriately.

Amino-acyl tRNA Synthetase

The Master Regulator Of The Proteome
By Jamal Ross

Abstract
Translation is the process by which functional proteins are made from mRNA transcripts. Protein synthesis is a crucial biological phenomenon for complex life, as it is responsible for making sense of the genetic code in order to produce cells which display unique structural and biochemical properties, and thus underpins the process of cell specialisation. The genetic code is like an instruction manual to make all of the bodies biological machinery. The expression of these genes leads to the downstream assembly of functional proteins that may either potentiate or inhibit specific pathways. Although gene expression is controlled primarily at the level of transcription, translational control is essential in order to ensure protein fidelity and prevent pathology. Amino-acyl tRNA synthetases (hereafter AARS) are responsible for loading amino acids onto their cognate tRNAs and must display a high level of specificity to be able to distinguish between structurally similar amino acids. In recent years, there has been a wealth of information to implicate AARS in pathological disease. This essay will focus on the most recent and illustrative discoveries about genetic mutations of AARS in pathology, on AARS regulating disease-related processes, and the potential use of AARS as pharmacological

targets and therapeutic reagents.

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The Process Of Translation: A Review Translation is the process of forming structural proteins from mRNA transcripts using ribosomes which are enormous complexes composed of three RNA molecules (5S, 16s, and 23S) and more than 50 proteins. The overall chemical reaction for translation is as follows:

The equivalent of two ATP are consumed in this reaction. The first is consumed by forming the

aminoacyl-tRNA ester linkage and the second is used to drive the reaction forward. tRNA therefore serves as the adapter molecule that binds to a specific codon and brings with it a specific amino acid for incorporation into the polypeptide chain. People often talk of translation as being composed of three main steps, namely: Initiation, elongation, and termination. However, there is a step that precedes all of these processes and is ,in actuality, the most important aspect of protein translation. This is known as the activation step and is crucial in maintaining protein fidelity. Activation is the process by which an amino acid is correctly loaded onto its cognate tRNA molecule. Without this highly regulated step, any downstream translation event would lead to an incorrectly folded protein. In 1998, evidence was provided for amino-acylation of tRNA in the nucleus of Xenopus laevis (African- clawed frog), which implies that AARS exist in the nucleus. Since these enzymes are too large to be imported by passive diffusion, they are thought to use sequence signatures to enter the nucleus specifically by nuclear

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Hell shake down the nations in a sieve of destruction, herd them into a dead end
Isaiah 30:27

localisation sequences and enter through importins. Intranuclear amino-acylation is, therefore, a proofreading mechanism to ensure that only correctly charged tRNAs are exported into the cytoplasm for translation ( Szymanski et al, 2000). That being said, aminioacylation events have also been known to occur within the cytoplasm. . So what is the big problem if a few mistranslated proteins are produced? After all, there would be a pool of correctly folded proteins to compensate for any mis-folded protein products and surely the ERAD pathway is able to degrade them. Well, the problem is that most functional components of the cell are proteins, therefore a mis-folded protein in the DNA replication machinery or in DNA repair mechanisms can lead to spontaneous mutagenic events. These mutagenic events would lead to hard-coded errors in the DNA which are subsequently propagated into all daughter cell lineages. This explains why AARS are implicated in pathologies relating to cancer, autoimmune conditions and neuropathies.

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Class I and Class II AARS

Jamal Ross

Different approaches to the same problem

AARS and their Molecular Structures Firstly, in order to understand function, we must appreciate structure. AARS are grouped into two classes, class I and class II, based on their mechanism of action. Class I AARS bind to tRNA from the minor groove and attach amino acids to the 2OH of adenosine (Figure 3B). They have a 5 handed parallel beta sheet motif known as a Rossman fold which is the site of aminoacylation. The aminoacylation site is composed of a unique ATP-binding consensus sequence HIGH-KMSKS. Class II AARS approach from the major groove and attach an amino
FIGURE 3

acid to the 3OH. They have

a 7 handed beta sheet with three alpha helices. If we look at figure 3, we can observe that the AARS for Ile, Val and Gln cradle the tRNA by the anticodon loop and the CCA arm interacts with the binding site in the top right; these are class I AARS. However, The AARS
FIGURE 3B

for Phe and Thr interact with a different face of tRNA and are class II AARS. These two enzyme classes provide solutions to the same problem but approach it by a different mechanism.

FIGURE 4

Whilst they differ in their exact mechanism of action, they do share a common evolutionary history. Both classes are thought to have been produced in pairs where they initially worked together in the prehistoric genome. However, they slowly evolved to develop their own unique functions without a dependence on each other (Fukai et al, 2000) .

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Class I and Class II AARS

Jamal Ross

Different approaches to the same problem

Figure 4 exemplifies the classification of AARS and the sub-classification into 6 classes. Perhaps, the most striking feature of AARS sub-classification is that specific pairings can occur. For example, class Ia AARS can bind to the same tRNA as a class IIa AARS without any sterical inhibition (Szymanski at al, 2000) . This is thought to occur because these enzymes had previously worked together and could bind to opposite faces of the same tRNA molecule. Therefore, class Ib enzymes could not bind to the same tRNA as IIc enzymes without sterically inhibiting each other. This is evidence of their common evolutionary origin and the existence of a common ancestor for the three kingdoms. The thing that distinguishes specific tRNA molecules from other tRNA molecules is its unique anticodon region. Mutagenesis studies, however, indicate that not all synthetases interact with their cognate tRNAs anticodons. E.Coli tRNAcys differs from tRNAala at 40 positions. It has a G-C bond at position 3:70. When mutated to the non-watson crick base pair G-U, the tRNAcys is recognised by alanyl tRNA synthetase as though it were tRNAala . This demonstrates that AARS can recognise tRNA by different positions in the microhelix structure and therefore, the integrity of its sequence may potentially create yet another area for pathology. Whether or not this is a physiologically occurring phenomenon is not known but it is likely to be the cause in some cases (Stryer, 2012). In essence, the RNA world theory (Bernhardt et al.,2012) proposes a theory for the evolution of all forms of life. It is thought that in the beginning there were only simple RNA molecules. Some of these molecules possessed a secondary structure and elicited enzymatic-like functions. These functions governed simple processes so only basic life forms could be supported. However, at some point there was a cross over into the formation of proteins. Proteins opened up a new domain to create complex structures with specific functions. These processes were able to form interactions and feedback mechanisms, all of which were necessary for developing higher organisms. Importantly, aminoacyl tRNA synthetases are the bridge between the RNA world and the protein world as we know today. They played a pivotal role in the process of evolution of all species in addition to the ribosomes which are an example where remnants of the RNA world (5S, 23S and 16S) possess catalytic function.

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The Double Sieve

The Gift Of Discernment
By Jamal Ross

The Double Sieve and The gift of discernment The potential for error in translation is exacerbated by the many steps and molecules involved. There is an ongoing conflict between two requirements: the rate of translation must be both accurate and yet fast enough to meet the demands of the cell. In table 1 we can see that the probability of forming a protein with no errors is dependent on how many amino acids it contains and the frequency of inserting an incorrect amino acid. Pauling estimated that if AARS depended on a one step amino-acylation pathway then L-valine would be mis-incorporated in place of L-Isoleucine , 20% of the time (Nureki et al, 2008).

(Stryer, 2012)

An value of around 10 is needed to maintain high fidelity in larger proteins that have 1000 amino acids. The human body has an error rate close to this value although it may be reduced for larger proteins by taking more time in the proofreading process. Therefore, the observed error rate of protein translation is approximately 200 times lower than it should be. This suggests that there is an underlying mechanism which ensures this high fidelity. AARS have two primary functions in translation: (i), activation of the amino acid with ATP and (ii) to charge the amino-acyl moiety to the terminal 3 adenosine of tRNA. In the amino-acylation reaction, amino acid selection is the least specific step. In figure 1, IleRS is able to bind both LValine and L-isoleucine in the first sieve but only L-valine can bind the second sieve in the editing domain and is subsequently hydrolysed. Amino acids that are larger than Ile are excluded at the first sieve. The first sieve which accommodates both L- isoleucine and L-valine is on the amino-acylation domain containing the Rossmann fold, whereas the second sieve, which is specific to L-valine, is found in a globular -barrel CP domain that protrudes from the aminoacylation domain.

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The Double Sieve

The Gift Of Discernment
By Jamal Ross

Isoleucine is a particularly difficult example. It is recognized by an isoleucine-shaped hole in the enzyme, which is too small to fit larger amino acids like methionine and phenylalanine, and too hydrophobic to bind anything with polar side-chains. But, the slightly smaller amino acid valine, different by only a single methyl group, also fits nicely into this pocket, binding instead of isoleucine in about 1 in 150 times. This is far too many
FIGURE 5 ; Nureki et al,

errors for living organisms, so corrective steps

must be taken to reduce this error rate. Isoleucyl-tRNA synthetase (PDB entry 1FFY) solves this problem with a second active site, which performs an editing reaction. Isoleucine does not fit into this site, but errant valine does. The mistake is then cleaved away, leaving the tRNA ready for a properly-placed Isoleucine amino acid. This proofreading step improves the overall error rate to about 1 in 3,000. Mutagenesis studies have revealed some of the underlying mechanisms by which AARS exhibit their specificity. The double sieve mechanism explains how AARS differentiate between chemically similar amino acids such as Valine and Isoleucine. Figure 6 demonstrates murine
Knowing the difference Scentists still marvel at the ability of AARS to discriminate between very chemically similar Amino acids. Probing into this mechanism may help in the future with designing articial enzymes for enzyme replacement therapy.

defective ValRS where there has been a T535P mutation. The graph on the left shows that this mutation does not affect the amino-acylation capabilities of the enzyme. However, the graph on the right shows that the mutation impairs the ability of the enzyme to clear Thr-tRNA-Val. This proves that there is a proofreading mechanism which screens the charged tRNA molecule and then cleaves a mis-incorporated amino acid.

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The Double Sieve

The Gift Of Discernment
By Jamal Ross

The first-sieve discriminates amino acids by size; any amino-acid that is smaller than the cavity will be able to rattle around in a smaller cavity. On the other hand, it is energetically unfavourable for a larger amino acid to be forced into a much smaller cavity. The second sieve is responsible for discriminating on the basis of hydrophobicity. An amino acid with fewer

FIGURE 6 (Schimmel et al, 2008)

hydrophobic contacts will be hydrolysed whereas the amino acid with greater hydrophobic contacts will remain bound to the tRNA by its ester linkage.

Figure 7

Image Reproduced from (Fukai et al., 2000)

Figure 7 represents Valine tRNA synthetase and demonstrates how this AARS must be able to discriminate between threonine and valine, which differ only by a single methyl group. Isoleucine, however, is naturally excluded from the first sieve by its large size. Threonine has fewer hydrophobic contacts than valine and binds to the second sieve where it is then cleaved by two peptides. If valine was conjugated with the tRNA then it remains attached to the tRNA to be incorporated into a growing polypeptide chain.

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Non-canonical Functions of AARS

Its More than just translation
By Jamal Ross

The canonical functions of AARSs, including aminoacylation and editing, are highly conserved throughout the three kingdoms. However, during evolution from prokaryotes to vertebrates, including mammals, certain AARSs acquired appended domains with unique structural characteristics that are neither a part of the enzymatic core nor present in bacterial homologues. These newly evolved domains, generally affixed to the amino or carboxy terminus, are not essential for tRNA charging, but instead are responsible for noncanonical activities unrelated to aminoacyla- tion (Guo et al, 2010), including translation control, transcription regulation, signal transduction, cell migration, angiogenesis, inflammation, and tumourigenesis. Emerging evidence suggests that defects in either canonical or noncanonical functions can cause or contribute to human diseases. Potential associations of AARS with several types of cancer through aberrant expression and undesired interactions have been described (Kim et al, 2011,

2012b). We now know that many of the idiopathic inflammatory myopathies are closely linked to mutations in AARS. The idea that AARS are only implicated in translation is a highly oversimplified view that is drastically changing due to emerging evidence in medical research. An interesting aspect is that AARS are implicated in cytokine-like activity and the processes involved in inflammatory pathways. Perhaps then, research in this field will reveal new ways to modulate the immune response and may even help in defending the body from retroviral infections (Boonyalai et al., 2011). AARS are also implicated in mTOR, IFN- and p53 signaling ( Guo et al, 2013). LysRS, SerRS and PheRS were shown to be responsible for the synthesis and turnover of diadenosine tetraphos- phate (Ap4A) that plays an important role in the response of bacterial and eukaryotic cells to a variety of stress conditions (Zamecnike et al, 1966) function within the cell as a new class of signalling mol- ecules, called alarmones ( Nishimura et al, 1998) . Furthermore, an EMAP II like domain in carboxy terminal of human TyrRS: Shows cytokine activity. Leukocyte and monocyte chemotaxis. (Wakasugi et al, 1999)

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AARS in Pathological Disease

The Clinical Implications
By Jamal Ross

AARS in pathological Disease At first glance, it may seem that AARS would not be implicated in serious pathological disease since a mistranslation of protein is at the proteomic level and not the genomic level, therefore any errors would not be propagated in DNA replication to daughter cells. These errors would seemingly be transient and functional proteins would still be produced alongside the abnormal proteins. However, as mistranslation affects proteins, it can also affect the DNA replication machinery, such as DNA polymerase and DNA ligase, all of which are proteins. This creates a situation where errors may now be hard-coded into the DNA causing a mutagenic event. In addition to this, repair enzymes are also affected. In a recent study by, aging colonies of E.coli were tested for the rate at which they underwent spontaneous mutation to yield Rifampicin resistance (Bayat et al, 2012). It was found that bacteria that possessed a defective editing mechanism were more likely to incur spontaneous mutation in their DNA and thus were more likely to develop rifampicin resistance. Furthermore, chaperones which are implicated in the repair of mis-folded proteins are themselves proteins. Therefore, they are subject to the same stresses as any other protein. If a mis-charged tRNA is used to form a chaperone protein then it could lead to a magnified downstream effect. If the master regulator of a process is abnormal then its restorative properties will no longer be effective. Because mutations in the site for editing do not disrupt the essential aminoacylation function, mild defects in editing can in principle be vertically transmitted in the population.

Autoimmmunity In 1962 Weigle carried out some experiments which may demonstrate how editing defects can elicit an autoimmune condition. Anthranilic acid was coupled to thyroglobulin and injected into rabbits. As predicted, the rabbit raised antibodies against the conjugated hapten of Anthranilic acid and thyroglobulin. However, over time, the rabbits also developed thyroiditis whereby antibodies were directed specifically against thyroglobulin alone. It is conceivable then, that when mistranslation occurs, immunologic protein variants could also trigger an autoimmune reaction to the native protein.

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AARS in Pathological Disease

The Clinical Implications
By Jamal Ross

AARS have been implicated in a wide range of diseases such as inflammatory myopathies. They are thought to be autoimmune disorders, in which the bodys white blood cells (that normally fight disease) attack blood vessels, normal muscle fibers, and connective tissue in organs, bones, and joints. Polymyositis is chronic inflammation of the muscles. Dermatomyositis is a connective-tissue disease related to polymyositis (PM) that is characterized by inflammation of the muscles and the skin. While Dermatomyositis most frequently affects the skin and muscles, it is a systemic disorder that may also affect the joints, the esophagus, the lungs, and, less commonly, the heart. Amyotrophic lateral sclerosis is a debilitating disease with varied etiology characterized by rapidly progressive weakness, muscle atrophy and fasciculations, muscle spasticity, difficulty speaking (dysarthria), difficulty swallowing (dysphagia), and difficulty breathing (dyspnea). Diseases which affect the functioning of mitochondria usually result in neuropathy and myopathy because nerve cells and muscle cells are the most energy-dependent cells in the human body. Therefore, it is no surprise that proteins which are either encoded in

FIGURE 7a

FIGURE 7b

FIGURE 7c

mitochondrial DNA or that interact with mitochondrial components are potential hotspots for these diseases. Figure 7a and 7c show a 45 year old patient with dermatomyositis and a sparse discolouration of the skin. FIgure 7b shows a histological section of a patient with polymyositis and the abnormal recruitment of white blood cells which is responsible for driving the destructive inflammatory crisis.

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Neurodegeneration In a recent forward genetic screen to identify genes required for neuronal function and survival in Drosophila melanogaster, it was subsequently found that mutated methionyl-tRNA synthetase (Homologue of human MARS2), led to degeneration of neuron photoreceptors (Bayat et al, 2012).

FIGURE 9

FIGURE 10

FIGURE 11

Furthermore, it was found that MARS2 is mutated in human Autosomal Recessive Spastic Ataxia Leukoencehalopathy (hereafter ARSAL) and displays a similar phenotype as observed in D.melanogaster. Figure 9 demonstrates the sequence homology between the drosophila Aats-met gene ( AARS for methionine) and the human homolog, MARS2. In Figure 10, mitochondria have been tagged with GFP whilst MARS2 has been tagged with FLAG. This experiment shows that they both co-localize in the mitochondria which demonstrates, with some certainty, that MARS2 is a mitochondrial protein. Figure 11 is a PCR amplification of a portion encoding for MARS2. In the individuals E9, E10 and E11 there is a 280bp deletion leading to ARSAL. This differs from individual C6 and C8 who show a 300bp insertion but also present with ARSAL.

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Figure 12 is a T2 weighted MRI that demonstrates some of the gross pathological changes that occur in Autosomal recessive spastic ataxia with leukoencephalopathy. These changes include cortical and cerebellar atrophy of white matter and thinning of the corpus callosum. Patient A has the most severe phenotype which is accompanied with the most changes in the white matter. Patient E has the least thinning of the corpus callosum and fewer symptoms of the disease. Figure 13 is a pedigree of 17 families in Quebec who were screened for ARSAL
FIGURE 12 (Thiffault et al, 2006)

mutations.

This revealed a recessive pattern of inheritance that predominantly occurs in a community in East-Central Canada due to a phenomenon called the founder effect. This occurs when a group of people who are not representative of the general population reproduce and , consequently, the prevalence of rare defective alleles increase in that community. Figure 13

Prior to this study, ARSAL remained amongst a heterogenous group of poorly classified neurological diseases. In a clinical context, it is not crucial to know the specific underlying etiology of each disease but rather to be able to manage the symptoms appropriately. However, in saying that, if we are to ever be innovators an not just assembly-line physicians then it becomes essential to know the underlying mechanism of a disease. In knowing this, it creates a potential avenue for new drug targets and perhaps, even finding a cure. Therefore, the emergence of defective AARS in the development of ARSAL will allow researchers to provide more targeted therapy to patients allowing them to have a better quality of life. In addition, a deeper understanding of AARS pathophysiology may give clues to our evolutionary origin and provide answers for many areas in synthetic biology.

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Biomimicry -
Emulating Natures genius
By Jamal Ross

Chang shan () is a herb that has been used for well over 2000 years in traditional chinese medicine in the treatment of malaria. Its popularity as a treatment has dwindled as the popular Artemisinin herb () has replaced much of its use. However, it has recently been discovered that Chang shan has a selective suppressive effect on the immune system. This property has led to its use in a range of autoimmune conditions such as rheumatoid arthritis, psoriasis and thyroiditis.

In the past week I went to five different traditional chinese clinics in Shenyang trying to find out more information about this herb and its clinical use. All of the doctors I spoke to were aware of its use in malaria but were not aware of its recent application to autoimmune diseases. It seems as though this information has not yet trickled back down into the traditional medical domain. Moreover, this herb is no longer being
(Zhou et al, 2013)

stocked by Chinese First or Second affiliated hospitals. Perhaps, this is because most are not

aware of its potent effect in treating autoimmune diseases and also the increasing acceptance of Western medicine amongst the chinese populace . In a recent study undertaken by Harvard university (Zhou et al, 2013), they found that the active component in Chang shan that suppresses the immune system is a compound called Halofuginone. It acts to specifically inhibit Th17 cells without dampening the immune systems response to

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invading pathogens. Th17 cells were only recently discovered in 2007 and have been implicated in driving autoimmune diseases by releasing pro-inflammatory cytokines. The way that this compound specifically inhibits Th17 cells is through blocking proline-aminoacyl tRNA synthetase. It does this by binding with ATP and mimics the effects of proline-deprivation. It is still unknown why proline is essential for Th17 maturation but not for other immune cells. This finding has led to a huge collaborative effort at Harvard university to reverse engineer the compound Halofuginone ,with the hope of gaining FDA approval. In one way, monetizing this drug and making it a marketable commodity is a sad re-occurring feature of todays medical field. Many people will not be able to benefit from this drugs therapeutic effect and some will never be made aware of its existence. Chang shan is just one example of many traditional chinese medicines that pharmaceutical companies have reverse engineered to create patented FDA approved medication. It could be argued that patenting and monetizing a compound which is already available on the public market for around 8 RMB is somewhat unethical. Nevertheless, the biochemical mechanisms involved suggest that AARS are viable drug targets for a whole range of pathological conditions.

FIGURE 13

When I first came to China I thought that traditional Chinese medicine (TCM) was a science but now I have learnt that it is not just a science but it is a philosophy. In Western civilization we often hear of the terms yin and yang to describe seemingly opposing forces in nature that really work harmoniously together. Similarly, a herb is really a concoction of thousands of compounds, some of which potentiate the effect of the main active compound. In Western medicine we often try to explain disease in terms of one drug, one target and we therefore try to isolate and purify the main active compound that elicits the desired therapeutic effect. However, diseases are not linear processes as there is a wealth of cross-talk between different signaling pathways and molecular interactions. Therefore, the holistic approach in TCM is more reflective of the diseased health state by targeting multiple drug targets at once. In recent years there has been an emerging trend towards this philosophy under the branch of science termed network pharmacology. Figure 13 shows a network of nodes which represent proteins and a highly connected node is called a hub. If a hub is defective, then , generally speaking, it will be more disastrous for the overall system than a node with fewer interactions. Working in China has given me a very unique opportunity to learn about the wonders of

TCM which are now gaining the attention of many researchers over the world. Personally, I believe it to be paramount to learn about these traditional remedies to inform patients of their therapeutic options, particularly those that may not be able to afford alternatives. Below, table 3 demonstrates some of the FDA approved drugs that have been derived from TCM remedies in recent

years.

(Zhou et al., 2013)

In the next decade I propose that modern antibiotics will be supplanted by newly developed pharmaceutical agents as resistance emerges in community and nosocomial infections. Pseudomonic acid (mupirocine) which inhibits specific AARS enzymes has been used as an alternative to vancomycin for bacterial infections. Phenylthiazole 1,3,5 traizine derivatives inhibit candida albicans leucyl-tRNA synthetases for a new class of antifungals (Singh et al, 2013). In comparison with, for example, terbinafine and itraconazole, compound C10 (AN2690) is a very promising candidate for treatment of ungual and periungual infections with improved nail penetration and low keratin binding. Furthermore, a recent comprehensive bioinformatic analysis revealed 26 AARS and five freestanding editing domains in Leishmaniasis major (protozoa). The unique features identified in this work provide rationale for designing inhibitors against parasite aminoacyl tRNA synthetases and their paralogs (Gowri et al , 2012). Lastly, AARS are being intensely studied as new drug targets for autoimmune conditions and cancer therapy (Zhou et al, 2013). We are at the beginning of an exciting new era of academic research and innovation, and although we are constantly presented with many obstacles, in that same domain lies a great opportunity to overcome.

REFERENCES

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References
AARS
By Jamal Ross

Michael Ibba and Dieter Soll (2000) Aminoacyl-tRNA Synthesis. Annual Review of Biochemistry 69, 617-650. John G. Arnez and Dino Moras (1997) Structural and Functional Considerations of the Aminoacylation Reaction. Trends in Biochemical Sciences 22, 211-216. Stephen Cusak (1995) Eleven Down and Nine to Go. Nature Structural Biology 2, 824-831. Jonathan J. Burbaum and Paul Schimmel (1991) Structural Relationships and the Classication of Aminoacyl-tRNA Synthetases. Journal of Biological Chemistry 266, 16965-16968. Udaya Pratap Singh, Hans Raj Bhat, Prashant Gahtori, and Ramendra K SingH (2013). Hybrid phenylthiazole and 1,3,5-triazine target cytosolic leucyl-tRNA synthetase for antifungal action as revealed by molecular docking studies. In Silico Pharmacology 2013, 1:3 V S Gowri, Indira Ghosh, Amit Sharma, and Rentala Madhubala. Unusual domain architecture of aminoacyl tRNA synthetases and their paralogs from Leishmania major. BMC Genomics 2012, 13:621
http://www.ninds.nih.gov/disorders Guo et al. Nature Chemical Biology 9, 145153 (2013) doi:10.1038/nchembio.1158 Sadakh et al. Aminoacyl-tRNA synthetase inhibitors as antimicrobial agents: a patent review from 2006 till present. December 2012, Vol. 22, No. 12 , Pages 1453-1465 (doi:10.1517/13543776.2012.732571) LI Shao*, ZHANG Bo .Traditional Chinese medicine network pharmacology: theory, methodology and application. Chinese Journal of Natural Medicines 2013, 11(2): 0110!0120 Zamecnik, P.C., Stephenson, M.L., Janeway, C.M. & Randerath, K. (1966) Enzymatic synthesis of diadenosine tetraphosphate and diadenosine triphosphate with a puried lysyl-tRNA synthetase. Biochem. Biophys. Res. Commun. Nishimura, A. (1998) The timing of cell divi- sion: Ap4A as a signal. Trends Biochem. Sci. 23, 157159.
Wakasugi, K. & Schimmel, P. (1999) Two dis- tinct cytokines released from a human amino- acyl-tRNA synthetase. Science 284, 147151.

Harpers Biochemistry. (2012) Arshan Nasir, Kyung Mo Kim, and Gustavo Caetano-Anolles. BMC Evolutionary Biology 2012, 12:1. doi: 10.1186/1471-2148-12-156 Valeria C Caballero, Viviana P Toledo, Cristian Maturana, Carolyn R Fisher, Shelley M Payne, and Juan Carlos Salazar.BMC Microbiology 2012, 12:1. doi:10.1186/1471-2180-12-226 Cheryl P Andam, Timothy J Harlow, R Thane Papke, and J Peter Gogarten. BMC Evolutionary Biology 2012, 12:1. doi:10.1186/1471-2148-12-85

Thiffault, I. (2006). A new autosomal recessive spastic ataxia associated with frequent white matter changes maps to 2q33-34. Brain, 129(9), 23322340. doi:10.1093/brain/awl110

Yao, P., & Fox, P. L. (2013). Aminoacyl-tRNA synthetases in medicine and disease. EMBO Molecular Medicine, 5(3), 332343. doi:10.1002/emmm.201100626 Aminoacyl-tRNA Synthetase and Its Application on In Vivo Incorporation of Non-natural Amino Acids into Proteins. (2003). Aminoacyl-tRNA Synthetase and Its Application on In Vivo Incorporation of Non-natural Amino Acids into Proteins, 168. Nasir, A., Kim, K. M., & Caetano-Anolles, G. (2012). Giant viruses coexisted with the cellular ancestors and represent a distinct supergroup along with superkingdoms Archaea, Bacteria and Eukarya. BMC Evolutionary Biology, 12(1), 11. BMC Evolutionary Biology. doi:10.1186/1471-2148-12-156 Mehta, S. Q., Hiesinger, P. R., Beronja, S., Zhai, R. G., Schulze, K. L., Verstreken, P., Cao, Y., et al. (2005). Mutations in Drosophila sec15 Reveal a Function in Neuronal Targeting for a Subset of Exocyst Components. Neuron, 46(2), 219232. doi:10.1016/j.neuron.2005.02.029 Zhou, X. L., Ruan, Z. R., Huang, Q., Tan, M., & Wang, E. D. (2012). Translational delity maintenance preventing Ser mis-incorporation at Thr codon in protein from eukaryote. Nucleic Acids Research, 41(1), 302314. doi:10.1093/nar/gks982 Zhou, H., Sun, L., Yang, X.-L., & Schimmel, P. (2012). ATP-directed capture of bioactive herbal-based medicine on human tRNA synthetase. Nature, 494(7435), 121124. doi:10.1038/nature11774 Bayat, V., Thiffault, I., Jaiswal, M., Ttreault, M., Donti, T., Sasarman, F., Bernard, G., et al. (2012). Mutations in the Mitochondrial Methionyl-tRNA Synthetase Cause a Neurodegenerative Phenotype in Flies and a Recessive Ataxia (ARSAL) in Humans. (D. R. Green, Ed.)PLoS Biology, 10(3), e1001288. doi:10.1371/journal.pbio.1001288.s014 Panwar, B., & Raghava, G. P. (2010). Prediction and classication of aminoacyl tRNA synthetases using PROSITE domains. BMC Genomics, 11(1), 507. doi:10.1186/1471-2164-11-507 Bernhardt, H. S. (2012). The RNA world hypothesis: the worst theory of the early evolution of life (except for all the others)(a). Biology direct, 7, 23. doi:10.1186/1745-6150-7-23 Meier, P., Schirmer, S. H., Lansky, A. J., Timmis, A., Pitt, B., & Seiler, C. (2013). The collateral circulation of the heart. BMC Medicine, 11(1), 143. doi:10.1186/1741-7015-11-143 Brewster, L. M., & Seedat, Y. K. (2013). Why do hypertensive patients of African ancestry respond better to calcium blockers and diuretics than to ACE inhibitors and beta-adrenergic blockers? A systematic review. BMC Medicine, 11(1), 141. doi:10.1186/1741-7015-11-141