Kontrola kvalitete pri biosintezi aminoacil-tRNA

The fidelity of translation is determined at two major points: the accuracy of aminoacyl-tRNA selection by the ribosomes and synthesis of cognate amino acid/tRNA pairs by aminoacyl-tRNA synthetases (aaRSs) in the course of the aminoacylation reaction. The most important point in aminoacylation is the accurate recognition of cognate substrates coupled with discrimination of non-cognates. While this is generally accomplished by a single enzyme, we have recently found that discrimination against lysine analogues requires the existence of two unrelated lysyl-tRNA synthetases. For other amino acids, initial recognition is not sufficiently accurate with errors being corrected by an intrinsic editing activity. Recent studies indicate how editing prevents the misinterpretation of phenylalanine as tyrosine in the genetic code and have shown the importance of this process in vivo. More recent studies indicate that while these editing reactions are critical in the cytoplasm, some are absent from mitochondria suggesting that the overall fidelity of protein synthesis might be reduced in this compartment.Vjernost translacije bitno ovisi o točnosti dvaju koraka: odabiru aminoacil-tRNA na ribosomu i sintezi ispravnih aminoacil-tRNA pomoću odgovarajućih aminoacil-tRNA-sintetaza u reakciji aminoaciliranja. Najvažniji događaj u aminoaciliranju precizno je prepoznavanje pripadnih supstrata (tRNA i aminokiseline) i diskriminacija nepripadnih. Iako taj posao uglavnom obavlja po jedan enzim za svaki par tRNA : aminokiselina, nedavno smo ustanovili da su za diskriminaciju analoga lizina potrebne dvije različite lizil-tRNA-sintetaze. U nekim drugim slučajevima otkriveno je da su pogreške u odabiru tRNA i njihovih pripadnih aminokiseline i suviše velike, pa je nužan naknadni popravak pogrešnih produkata u reakciji aminoaciliranja, koji također mogu katalizirati neke aminoacil-tRNA-sintetaze. Na primjeru krivog odabira tirozina umjesto fenilalanina, te naknadnog popravka, pokazano je kako je mogućnost korekcije važna u sprečavanju pogrešne translacije genetičkog koda in vivo. Najnovija istraživanja pokazala su da su mehanizmi popravka od ključne važnosti u citoplazmi, no neki se ne zbivaju u mitohondriju, ukazujući na smanjenu ukupnu točnost biosinteze proteina u ovom staničnom odjeljku

FARS2 mutations presenting with pure spastic paraplegia and lesions of the dentate nuclei.

Mutations in FARS2, the gene encoding the mitochondrial phenylalanine-tRNA synthetase (mtPheRS), have been linked to a range of phenotypes including epileptic encephalopathy, developmental delay, and motor dysfunction. We report a 9-year-old boy with novel compound heterozygous variants of FARS2, presenting with a pure spastic paraplegia syndrome associated with bilateral signal abnormalities in the dentate nuclei. Exome sequencing identified a paternal nonsense variant (Q216X) lacking the catalytic core and anticodon-binding regions, and a maternal missense variant (P136H) possessing partial enzymatic activity. This case confirms and expands the phenotype related to FARS2 mutations with regards to clinical presentation and neuroimaging findings

Effects of training on postural control and agility when wearing socks of different compression levels

Study aim: The aim of this study was to evaluate the effects of training while wearing socks differing in compression level (clinical, sub-clinical, regular) on performance of static and dynamic balancing and agility tasks in healthy, physically active people. We sought to understand whether socks with different compression properties supported postural regulation and agility task performance by enhancing somatosensory perception, unskewed by specific age range effects. Material and methods: Participants comprised 61 adults aged 18-75 years, divided into three groups (two experimental groups wearing clinical or sub-clinical level compression socks, and one control group wearing regular non-compression socks during training). An 8-week (2 × 1h per week) intervention programme was administered to train static and dynamic balance and postural control, leg strength and agility. Results: A mixed model ANOVA revealed no differences in static and dynamic balance and postural control and agility performance between clinical, sub-clinical, and control groups before and after training. All groups significantly improved their test performance, suggesting that training had some benefit on motor performance. Conclusions: These results raised interesting questions requiring further investigation to examine the effects of wearing socks (with and without different levels of compression) on motor behaviours in specific groups of elderly vs. young participants, in physically active vs. less physically active people, and in performance settings outside standardized laboratory tests to study applications in natural performance environments

Chronic Intermittent Ethanol Regulates Hippocampal GABA(A) Receptor Delta Subunit Gene Expression.

Chronic ethanol consumption causes structural and functional reorganization in the hippocampus and induces alterations in the gene expression of gamma-aminobutyric acid type A receptors (GABAARs). Distinct forced intermittent exposure models have been used previously to investigate changes in GABAAR expression, with contrasting results. Here, we used repeated cycles of a Chronic Intermittent Ethanol paradigm to examine the relationship between voluntary, dependence-associated ethanol consumption, and GABAAR gene expression in mouse hippocampus. Adult male C57BL/6J mice were exposed to four 16-h ethanol vapor (or air) cycles in inhalation chambers alternated with limited-access two-bottle choice between ethanol (15%) and water consumption. The mice exposed to ethanol vapor showed significant increases in ethanol consumption compared to their air-matched controls. GABAAR alpha4 and delta subunit gene expression were measured by qRT-PCR at different stages. There were significant changes in GABAAR delta subunit transcript levels at different time points in ethanol-vapor exposed mice, while the alpha4 subunit levels remained unchanged. Correlated concurrent blood ethanol concentrations suggested that GABAAR delta subunit mRNA levels fluctuate depending on ethanol intoxication, dependence, and withdrawal state. Using a vapor-based Chronic Intermittent Ethanol procedure with combined two-bottle choice consumption, we corroborated previous evidences showing that discontinuous ethanol exposure affects GABAAR delta subunit expression but we did not observe changes in alpha4 subunit. These findings indicate that hippocampal GABAAR delta subunit expression changes transiently over the course of a Chronic Intermittent Ethanol paradigm associated with voluntary intake, in response to ethanol-mediated disturbance of GABAergic neurotransmission

Cell-selective metabolic labeling of proteins

Metabolic labeling of proteins with the methionine surrogate azidonorleucine can be targeted exclusively to specified cells through expression of a mutant methionyl-tRNA synthetase (MetRS). In complex cellular mixtures, proteins made in cells that express the mutant synthetase can be tagged with affinity reagents (for detection or enrichment) or fluorescent dyes (for imaging). Proteins made in cells that do not express the mutant synthetase are neither labeled nor detected

Quality Control During Aminoacyl-tRNA Synthesis

The fidelity of translation is determined at two major points: the accuracy of aminoacyl-tRNA selection by the ribosomes and synthesis of cognate amino acid/tRNA pairs by aminoacyl-tRNA synthetases (aaRSs) in the course of the aminoacylation reaction. The most important point in aminoacylation is the accurate recognition of cognate substrates coupled with discrimination of non-cognates. While this is generally accomplished by a single enzyme, we have recently found that discrimination against lysine analogues requires the existence of two unrelated lysyl-tRNA synthetases. For other amino acids, initial recognition is not sufficiently accurate with errors being corrected by an intrinsic editing activity. Recent studies indicate how editing prevents the misinterpretation of phenylalanine as tyrosine in the genetic code and have shown the importance of this process in vivo . More recent studies indicate that while these editing reactions are critical in the cytoplasm, some are absent from mitochondria suggesting that the overall idelity of protein synthesis might be reduced in this compartment

Measurement of the Bottom-Strange Meson Mixing Phase in the Full CDF Data Set

We report a measurement of the bottom-strange meson mixing phase \beta_s using the time evolution of B0_s -> J/\psi (->\mu+\mu-) \phi (-> K+ K-) decays in which the quark-flavor content of the bottom-strange meson is identified at production. This measurement uses the full data set of proton-antiproton collisions at sqrt(s)= 1.96 TeV collected by the Collider Detector experiment at the Fermilab Tevatron, corresponding to 9.6 fb-1 of integrated luminosity. We report confidence regions in the two-dimensional space of \beta_s and the B0_s decay-width difference \Delta\Gamma_s, and measure \beta_s in [-\pi/2, -1.51] U [-0.06, 0.30] U [1.26, \pi/2] at the 68% confidence level, in agreement with the standard model expectation. Assuming the standard model value of \beta_s, we also determine \Delta\Gamma_s = 0.068 +- 0.026 (stat) +- 0.009 (syst) ps-1 and the mean B0_s lifetime, \tau_s = 1.528 +- 0.019 (stat) +- 0.009 (syst) ps, which are consistent and competitive with determinations by other experiments.Comment: 8 pages, 2 figures, Phys. Rev. Lett 109, 171802 (2012