Mistranslating the genetic code with leucine in yeast and mammalian cells.

Translation fidelity relies on accurate aminoacylation of transfer RNAs (tRNAs) by aminoacyl-tRNA synthetases (AARSs). AARSs specific for alanine (Ala), leucine (Leu), serine, and pyrrolysine do not recognize the anticodon bases. Single nucleotide anticodon variants in their cognate tRNAs can lead to mistranslation. Human genomes include both rare and more common mistranslating tRNA variants. We investigated three rare human tRNALeu variants that mis-incorporate Leu at phenylalanine or tryptophan codons. Expression of each tRNALeu anticodon variant in neuroblastoma cells caused defects in fluorescent protein production without significantly increased cytotoxicity under normal conditions or in the context of proteasome inhibition. Using tRNA sequencing and mass spectrometry we confirmed that each tRNALeu variant was expressed and generated mistranslation with Leu. To probe the flexibility of the entire genetic code towards Leu mis-incorporation, we created 64 yeast strains to express all possible tRNALeu anticodon variants in a doxycycline-inducible system. While some variants showed mild or no growth defects, many anticodon variants, enriched with G/C at positions 35 and 36, including those replacing Leu for proline, arginine, alanine, or glycine, caused dramatic reductions in growth. Differential phenotypic defects were observed for tRNALeu mutants with synonymous anticodons and for different tRNALeu isoacceptors with the same anticodon. A comparison to tRNAAla anticodon variants demonstrates that Ala mis-incorporation is more tolerable than Leu at nearly every codon. The data show that the nature of the amino acid substitution, the tRNA gene, and the anticodon are each important factors that influence the ability of cells to tolerate mistranslating tRNAs.

Anticodon sequence determines the impact of mistranslating tRNAAla variants.

Transfer RNAs (tRNAs) maintain translation fidelity through accurate charging by their cognate aminoacyl-tRNA synthetase and codon:anticodon base pairing with the mRNA at the ribosome. Mistranslation occurs when an amino acid not specified by the genetic message is incorporated into proteins and has applications in biotechnology, therapeutics and is relevant to disease. Since the alanyl-tRNA synthetase uniquely recognizes a G3:U70 base pair in tRNAAla and the anticodon plays no role in charging, tRNAAla variants with anticodon mutations have the potential to mis-incorporate alanine. Here, we characterize the impact of the 60 non-alanine tRNAAla anticodon variants on the growth of Saccharomyces cerevisiae. Overall, 36 tRNAAla anticodon variants decreased growth in single- or multi-copy. Mass spectrometry analysis of the cellular proteome revealed that 52 of 57 anticodon variants, not decoding alanine or stop codons, induced mistranslation when on single-copy plasmids. Variants with G/C-rich anticodons resulted in larger growth deficits than A/U-rich variants. In most instances, synonymous anticodon variants impact growth differently, with anticodons containing U at base 34 being the least impactful. For anticodons generating the same amino acid substitution, reduced growth generally correlated with the abundance of detected mistranslation events. Differences in decoding specificity, even between synonymous anticodons, resulted in each tRNAAla variant mistranslating unique sets of peptides and proteins. We suggest that these differences in decoding specificity are also important in determining the impact of tRNAAla anticodon variants.

Do multimodal search cues help or hinder teleoperated search and rescue missions?

In search and rescue missions, teleoperated rovers equipped with sensor technology are deployed into harsh environments to search for targets. To support the search task, unimodal/multimodal cues can be presented via visual, acoustic and/or haptic channels. However, human operators often perform the search task in parallel with the driving task, which can cause interference of attentional resources based on multiple resource theory. Navigating corners can be a particularly challenging aspect of remote driving, as described with the Cornering Law. Therefore, search cues should not interfere with cornering. The present research explores how unimodal/multimodal search cues affect cornering performance, with typical communication delays of 50 ms and 500 ms. One-hundred thirty-one participants, distributed into two delay groups, performed a target search task with unimodal/multimodal search cues. Search cues did not interfere with cornering performance with 50 ms delays. For 500 ms delays, search cues presented via the haptic channel significantly interfered with the driving task. Practitioner summary: Teleoperated rovers can support search and rescue missions. Search cues may assist the human operator, but they may also interfere with the task of driving. The study examined interference of unimodal and multimodal search cues. Haptic cues should not be implemented for systems with a delay of 500 ms or more.

Why cumulative loading calculated using non-weighted integration may not be suitable for assessing physical stress of the lower back: an empirical investigation of strain during lifting and lowering tasks.

When work-related physical stress is assessed using non-weighted integration, it is assumed that different loading conditions have a sufficiently comparable effect on the human body as long as the area under the loading curve is the same. Growing evidence cast doubt on whether this simple calculation can adequately estimate physical work-related strain. This study investigates in vivo, focussing on the lower back, whether the non-weighted method adequately reflects work-related physical strain of the lower back. Strain data resulting from lifting/lowering tasks performed in a laboratory study with an identical area under the loading curve but different load intensities were compared. Results showed that the non-weighted method does not sufficiently reflect the resulting muscular, cardiovascular and perceived strain but underestimates the influence of higher load intensity even in the range of medium physical exposure. Further research is needed regarding the determination of weighting factors and limit values. Practitioner Summary Given the dynamic nature of most physical work activities, the assessment of time-varying loading of the lower back is of particular interest in practice. Results show that the widely used non-weighted calculation method does not accurately reflect the resulting physical strain but underestimates the influence of higher load intensity.Abbreviations: MSD: musculoskeletal disorders; WMSD: work-related musculoskeletal disorders; KIM-LHC: Key Indicator Method Lifting, Holding, Carrying; RES: right erector spinae longissimus; LES: left erector spinae longissimus; HR: heart rate; RPE: rating of perceived exertion; EMG: surface electromyography; ECG: electrocardiography; SENIAM: Surface ElectroMyoGraphy for the Non-Invasive Assessment of Muscles; MVC: maximum voluntary contraction; ANOVA: analysis of variance; Std. error: standard error HIGHLIGHTSResults of this empirical investigation suggest that the widely used non-weighted calculation method is not fully suitable for calculating cumulative loading of the lower back.Even in the range of medium physical exposure the non-weighted calculation method does not accurately reflect the resulting strain on the human body but tends to underestimate the influence of higher load intensity due to higher external weight.Despite the same cumulative loading value obtained when using the non-weighted method, the resulting physical strain values are generally about 20-25% higher.The results may be used to further develop ergonomic assessment methods in order to avoid a misclassification of loading conditions and to prevent the risk of overexertion.

Sfp1 links TORC1 and cell growth regulation to the yeast SAGA-complex component Tra1 in response to polyQ proteotoxicity.

Chromatin remodeling regulates gene expression in response to the accumulation of misfolded polyQ proteins associated with Huntington's disease (HD). Tra1 is an essential component of both the SAGA/SLIK and NuA4 transcription co-activator complexes and is linked to multiple cellular processes, including protein trafficking and signaling pathways associated with misfolded protein stress. Cells with compromised Tra1 activity display phenotypes distinct from deletions encoding components of the SAGA and NuA4 complexes, indicating a potentially unique regulatory role of Tra1 in the cellular response to protein misfolding. Here, we employed a yeast model to define how the expression of toxic polyQ expansion proteins affects Tra1 expression and function. Expression of expanded polyQ proteins mimics deletion of SAGA/NuA4 components and results in growth defects under stress conditions. Moreover, deleting genes encoding SAGA and, to a lesser extent, NuA4 components exacerbates polyQ toxicity. Also, cells carrying a mutant Tra1 allele displayed increased sensitivity to polyQ toxicity. Interestingly, expression of polyQ proteins upregulated the expression of TRA1 and other genes encoding SAGA components, revealing a feedback mechanism aimed at maintaining Tra1 and SAGA functional integrity. Moreover, deleting the TORC1 (Target of Rapamycin) effector SFP1 abolished upregulation of TRA1 upon expression of polyQ proteins. While Sfp1 is known to adjust ribosome biogenesis and cell size in response to stress, we identified a new role for Sfp1 in the control of TRA1 expression, linking TORC1 and cell growth regulation to the SAGA acetyltransferase complex during misfolded protein stress.

Transfer RNAs: diversity in form and function.

As the adaptor that decodes mRNA sequence into protein, the basic aspects of tRNA structure and function are central to all studies of biology. Yet the complexities of their properties and cellular roles go beyond the view of tRNAs as static participants in protein synthesis. Detailed analyses through more than 60 years of study have revealed tRNAs to be a fascinatingly diverse group of molecules in form and function, impacting cell biology, physiology, disease and synthetic biology. This review analyzes tRNA structure, biosynthesis and function, and includes topics that demonstrate their diversity and growing importance.

Phospho-dependent recruitment of the yeast NuA4 acetyltransferase complex by MRX at DNA breaks regulates RPA dynamics during resection.

The KAT5 (Tip60/Esa1) histone acetyltransferase is part of NuA4, a large multifunctional complex highly conserved from yeast to mammals that targets lysines on H4 and H2A (X/Z) tails for acetylation. It is essential for cell viability, being a key regulator of gene expression, cell proliferation, and stem cell renewal and an important factor for genome stability. The NuA4 complex is directly recruited near DNA double-strand breaks (DSBs) to facilitate repair, in part through local chromatin modification and interplay with 53BP1 during the DNA damage response. While NuA4 is detected early after appearance of the lesion, its precise mechanism of recruitment remains to be defined. Here, we report a stepwise recruitment of yeast NuA4 to DSBs first by a DNA damage-induced phosphorylation-dependent interaction with the Xrs2 subunit of the Mre11-Rad50-Xrs2 (MRX) complex bound to DNA ends. This is followed by a DNA resection-dependent spreading of NuA4 on each side of the break along with the ssDNA-binding replication protein A (RPA). Finally, we show that NuA4 can acetylate RPA and regulate the dynamics of its binding to DNA, hence targeting locally both histone and nonhistone proteins for lysine acetylation to coordinate repair.

Pathways to disease from natural variations in human cytoplasmic tRNAs.

Perfectly accurate translation of mRNA into protein is not a prerequisite for life. Resulting from errors in protein synthesis, mistranslation occurs in all cells, including human cells. The human genome encodes >600 tRNA genes, providing both the raw material for genetic variation and a buffer to ensure that resulting translation errors occur at tolerable levels. On the basis of data from the 1000 Genomes Project, we highlight the unanticipated prevalence of mistranslating tRNA variants in the human population and review studies on synthetic and natural tRNA mutations that cause mistranslation or de-regulate protein synthesis. Although mitochondrial tRNA variants are well known to drive human diseases, including developmental disorders, few studies have revealed a role for human cytoplasmic tRNA mutants in disease. In the context of the unexpectedly large number of tRNA variants in the human population, the emerging literature suggests that human diseases may be affected by natural tRNA variants that cause mistranslation or de-regulate tRNA expression and nucleotide modification. This review highlights examples relevant to genetic disorders, cancer, and neurodegeneration in which cytoplasmic tRNA variants directly cause or exacerbate disease and disease-linked phenotypes in cells, animal models, and humans. In the near future, tRNAs may be recognized as useful genetic markers to predict the onset or severity of human disease.

Mistranslating tRNA identifies a deleterious S213P mutation in the Saccharomyces cerevisiaeeco1-1 allele.

Mistranslation occurs when an amino acid not specified by the standard genetic code is incorporated during translation. Since the ribosome does not read the amino acid, tRNA variants aminoacylated with a non-cognate amino acid or containing a non-cognate anticodon dramatically increase the frequency of mistranslation. In a systematic genetic analysis, we identified a suppression interaction between tRNASerUGG, G26A, which mistranslates proline codons by inserting serine, and eco1-1, a temperature sensitive allele of the gene encoding an acetyltransferase required for sister chromatid cohesion. The suppression was partial, with a tRNA that inserts alanine at proline codons and not apparent for a tRNA that inserts serine at arginine codons. Sequencing of the eco1-1 allele revealed a mutation that would convert the highly conserved serine 213 within ß7 of the GCN5-related N-acetyltransferase core to proline. Mutation of P213 in eco1-1 back to the wild-type serine restored the function of the enzyme at elevated temperatures. Our results indicate the utility of mistranslating tRNA variants to identify functionally relevant mutations and identify eco1 as a reporter for mistranslation. We propose that mistranslation could be used as a tool to treat genetic disease.

AMICAI: A Method Based on Risk Analysis to Integrate Responsible Research and Innovation into the Work of Research and Innovation Practitioners.

The integration of ethics into the day-to-day work of research and innovation (R&I) is an important but difficult challenge. However, with the Aachen method for identification, classification and risk analysis of innovation-based problems (AMICAI) an approach from an engineering perspective is presented that enables the integration of ethical, legal and social implications into the day-to-day work of R&I practitioners. AMICAI appears in particular capable of providing a procedural guidance for R&I practitioners based on a method established in engineering science, breaking down the object of consideration into partial aspects and prioritizing the innovation-based problems in dependence of potential risk. This enables the user to apply AMICAI continuously during all stages of the research and development (R&D) process and to analyze and choose between certain sociotechnical alternatives. In this way, problems that affect ethical, legal, and social aspects can be understood, reflected and considered in the mostly technically focused R&D process. The paper gives a general guidance about AMICAI by describing principles and assumptions, providing the steps of analysis and application aids, giving an example application, explaining the necessary adjustments of AMICAI compared to the methodical basis of failure mode, effects, and criticality analysis and discussing the advantages and limits. AMICAI's simple applications can stimulate interdisciplinary cooperation in the R&D process and be a starting point for the development of an "open RRI risk analysis platform" allowing society to evaluate innovation-based problems.

Prediction model of the effect of postural interactions on muscular activity and perceived exertion.

Musculoskeletal disorders are a prevalent disease in many Western countries. While a large number of ergonomic analyses and assessment methods are nowadays available, most current methods that assess exposure calculate overall risk scores of individual body segments without considering interaction effects of exposure variables. Therefore, a study was conducted that aimed at investigating and quantifying interaction effects of trunk inclination and arm lifting on ratings of perceived exertion (RPE) and muscle activity. A multiple regression model to predict musculoskeletal load under consideration of interaction effects was derived. The study revealed that there is a significant interaction effect of trunk inclination and arm lifting. Furthermore, final regression models explained variance in exposure variables in a range of R2 = 0.68 to R2 = 0.147 with a subset of two to three inputs. The predicative equations support the computer-based post-processing of sensor data. Practitioner summary: This article elaborates on the importance of interaction effects of working postures on assessment results of load. In practise, easy to-use-methods for an assessment of working postures are needed. Therefore, a regression model is derived, which facilitates the quantification of work load under consideration of interaction effects. The use of this regression model for the assessment of posture data gathered by range sensors is recommended. Abbreviations: RPE: rating of perceived exertion; MSD: musculoskeletal disorder; OWAS: ovako working posture analysing system; RULA: rapid upper limb assessment; LUBA: postural loading on the upper body assessment; REBA: rapid entire body assessment; OCRA: occupational repetitive action;S D: standard deviation; EMG: surface electromyography; LUT: left upper trapezius pars descendens; RUT: right upper trapezius pars descendens; LLT: left trapezius pars ascendens; RLT: right trapezius pars ascendens; LAD: left anterior deltoideus; RAD: right anterior deltoideus; LES: left erector spinae longissimus; RES: right erector spinae longissimus; SENIAM: surface electroMyoGraphy for the non-invasive assessment of muscles; MVC: maximum voluntary contraction; MANOVA: multivariate analysis of variance; ANOVA: analysis of variance; OLS: ordinary least squares; MANCOVA: multivariate analysis of covariance.

Targeted sequencing reveals expanded genetic diversity of human transfer RNAs.

Transfer RNAs are required to translate genetic information into proteins as well as regulate other cellular processes. Nucleotide changes in tRNAs can result in loss or gain of function that impact the composition and fidelity of the proteome. Despite links between tRNA variation and disease, the importance of cytoplasmic tRNA variation has been overlooked. Using a custom capture panel, we sequenced 605 human tRNA-encoding genes from 84 individuals. We developed a bioinformatic pipeline that allows more accurate tRNA read mapping and identifies multiple polymorphisms occurring within the same variant. Our analysis identified 522 unique tRNA-encoding sequences that differed from the reference genome from 84 individuals. Each individual had ~66 tRNA variants including nine variants found in less than 5% of our sample group. Variants were identified throughout the tRNA structure with 17% predicted to enhance function. Eighteen anticodon mutants were identified including potentially mistranslating tRNAs; e.g., a tRNASer that decodes Phe codons. Similar engineered tRNA variants were previously shown to inhibit cell growth, increase apoptosis and induce the unfolded protein response in mammalian cell cultures and chick embryos. Our analysis shows that human tRNA variation has been underestimated. We conclude that the large number of tRNA genes provides a buffer enabling the emergence of variants, some of which could contribute to disease.

Genetic selection for mistranslation rescues a defective co-chaperone in yeast.

Despite the general requirement for translation fidelity, mistranslation can be an adaptive response. We selected spontaneous second site mutations that suppress the stress sensitivity caused by a Saccharomyces cerevisiae tti2 allele with a Leu to Pro mutation at residue 187, identifying a single nucleotide mutation at the same position (C70U) in four tRNAProUGG genes. Linkage analysis and suppression by SUF9G3:U70 expressed from a centromeric plasmid confirmed the causative nature of the suppressor mutation. Since the mutation incorporates the G3:U70 identity element for alanyl-tRNA synthetase into tRNAPro, we hypothesized that suppression results from mistranslation of Pro187 in Tti2L187P as Ala. A strain expressing Tti2L187A was not stress sensitive. In vitro, tRNAProUGG (C70U) was mis-aminoacylated with alanine by alanyl-tRNA synthetase, but was not a substrate for prolyl-tRNA synthetase. Mass spectrometry from protein expressed in vivo and a novel GFP reporter for mistranslation confirmed substitution of alanine for proline at a rate of ∼6%. Mistranslating cells expressing SUF9G3:U70 induce a partial heat shock response but grow nearly identically to wild-type. Introducing the same G3:U70 mutation in SUF2 (tRNAProAGG) suppressed a second tti2 allele (tti2L50P). We have thus identified a strategy that allows mistranslation to suppress deleterious missense Pro mutations in Tti2.

Visualizing tRNA-dependent mistranslation in human cells.

High-fidelity translation and a strictly accurate proteome were originally assumed as essential to life and cellular viability. Yet recent studies in bacteria and eukaryotic model organisms suggest that proteome-wide mistranslation can provide selective advantages and is tolerated in the cell at higher levels than previously thought (one error in 6.9 × 10-4 in yeast) with a limited impact on phenotype. Previously, we selected a tRNAPro containing a single mutation that induces mistranslation with alanine at proline codons in yeast. Yeast tolerate the mistranslation by inducing a heat-shock response and through the action of the proteasome. Here we found a homologous human tRNAPro (G3:U70) mutant that is not aminoacylated with proline, but is an efficient alanine acceptor. In live human cells, we visualized mistranslation using a green fluorescent protein reporter that fluoresces in response to mistranslation at proline codons. In agreement with measurements in yeast, quantitation based on the GFP reporter suggested a mistranslation rate of up to 2-5% in HEK 293 cells. Our findings suggest a stress-dependent phenomenon where mistranslation levels increased during nutrient starvation. Human cells did not mount a detectable heat-shock response and tolerated this level of mistranslation without apparent impact on cell viability. Because humans encode ∼600 tRNA genes and the natural population has greater tRNA sequence diversity than previously appreciated, our data also demonstrate a cell-based screen with the potential to elucidate mutations in tRNAs that may contribute to or alleviate disease.

Mistranslation: from adaptations to applications.

BACKGROUND: The conservation of the genetic code indicates that there was a single origin, but like all genetic material, the cell's interpretation of the code is subject to evolutionary pressure. Single nucleotide variations in tRNA sequences can modulate codon assignments by altering codon-anticodon pairing or tRNA charging. Either can increase translation errors and even change the code. The frozen accident hypothesis argued that changes to the code would destabilize the proteome and reduce fitness. In studies of model organisms, mistranslation often acts as an adaptive response. These studies reveal evolutionary conserved mechanisms to maintain proteostasis even during high rates of mistranslation. SCOPE OF REVIEW: This review discusses the evolutionary basis of altered genetic codes, how mistranslation is identified, and how deviations to the genetic code are exploited. We revisit early discoveries of genetic code deviations and provide examples of adaptive mistranslation events in nature. Lastly, we highlight innovations in synthetic biology to expand the genetic code. MAJOR CONCLUSIONS: The genetic code is still evolving. Mistranslation increases proteomic diversity that enables cells to survive stress conditions or suppress a deleterious allele. Genetic code variants have been identified by genome and metagenome sequence analyses, suppressor genetics, and biochemical characterization. GENERAL SIGNIFICANCE: Understanding the mechanisms of translation and genetic code deviations enables the design of new codes to produce novel proteins. Engineering the translation machinery and expanding the genetic code to incorporate non-canonical amino acids are valuable tools in synthetic biology that are impacting biomedical research. This article is part of a Special Issue entitled "Biochemistry of Synthetic Biology - Recent Developments" Guest Editor: Dr. Ilka Heinemann and Dr. Patrick O'Donoghue.

Pin1: Intimate involvement with the regulatory protein kinase networks in the global phosphorylation landscape.

BACKGROUND: Protein phosphorylation is a universal regulatory mechanism that involves an extensive network of protein kinases. The discovery of the phosphorylation-dependent peptidyl-prolyl isomerase Pin1 added an additional layer of complexity to these regulatory networks. SCOPE OF REVIEW: We have evaluated interactions between Pin1 and the regulatory kinome and proline-dependent phosphoproteome taking into consideration findings from targeted studies as well as data that has emerged from systematic phosphoproteomic workflows and from curated protein interaction databases. MAJOR CONCLUSIONS: The relationship between Pin1 and the regulatory protein kinase networks is not restricted simply to the recognition of proteins that are substrates for proline-directed kinases. In this respect, Pin1 itself is phosphorylated in cells by protein kinases that modulate its functional properties. Furthermore, the phosphorylation-dependent targets of Pin1 include a number of protein kinases as well as other enzymes such as phosphatases and regulatory subunits of kinases that modulate the actions of protein kinases. GENERAL SIGNIFICANCE: As a result of its interactions with numerous protein kinases and their substrates, as well as itself being a target for phosphorylation, Pin1 has an intricate relationship with the regulatory protein kinase and phosphoproteomic networks that orchestrate complex cellular processes and respond to environmental cues. This article is part of a Special Issue entitled Proline-directed Foldases: Cell Signaling Catalysts and Drug Targets.

Peroxide-mediated oxidation and inhibition of the peptidyl-prolyl isomerase Pin1.

Pin1 is a phosphorylation-dependent peptidyl-prolyl isomerase that plays a critical role in mediating protein conformational changes involved in signaling processes related to cell cycle control. Pin1 has also been implicated as being neuroprotective in aging-related neurodegenerative disorders including Alzheimer's disease where Pin1 activity is diminished. Notably, recent proteomic analysis of brain samples from patients with mild cognitive impairment revealed that Pin1 is oxidized and also displays reduced activity. Since the Pin1 active site contains a functionally critical cysteine residue (Cys113) with a low predicted pK(a), we hypothesized that Cys113 is sensitive to oxidation. Consistent with this hypothesis, we observed that treatment of Pin1 with hydrogen peroxide results in a 32Da mass increase, likely resulting from the oxidation of Cys113 to sulfinic acid (Cys-SO(2)H). This modification results in loss of peptidyl-prolyl isomerase activity. Notably, Pin1 with Cys113 substituted by aspartic acid retains activity and is no longer sensitive to oxidation. Structural studies by X-ray crystallography revealed increased electron density surrounding Cys113 following hydrogen peroxide treatment. At lower concentrations of hydrogen peroxide, oxidative inhibition of Pin1 can be partially reversed by treatment with dithiothreitol, suggesting that oxidation could be a reversible modification with a regulatory role. We conclude that the loss of Pin1 activity upon oxidation results from oxidative modification of the Cys113 sulfhydryl to sulfenic (Cys-SOH) or sulfinic acid (Cys-SO(2)H). Given the involvement of Pin1 in pathological processes related to neurodegenerative diseases and to cancer, these findings could have implications for the prevention or treatment of disease.

Work interruptions of office workers: The influence of the complexity of primary work tasks on the perception of interruptions.

BACKGROUND: Research demonstrates that work interruptions are considered one of the most common work stressors. Understanding the mechanisms of work interruptions is therefore vital to reducing worker stress and maintaining performance. OBJECTIVE: The aim of this research is to investigate the influence of the frequency of work interruptions on subjective workload in the context of office work. Specifically, the mediating influence of interruption perception as well as the moderating influence of the complexity of the primary task are examined. METHOD: The work interruptions of 492 office workers in Germany were collected by means of a one-day diary study. A mediation model and a conditional indirect effect model were calculated to examine the influence of interruption frequency on subjective workload, mediated by the individual perception of these interruptions as well as moderated by the complexity of the primary work tasks. RESULTS: The analyses indicated a significant mediation and moderation. This implies that, on the one hand, the perception of work interruptions significantly mediates the relationship between the frequency of work interruptions and subjective workload. On the other hand, more complex primary work tasks seem to strengthen the positive relationship between interruption frequency and perceived interruption overload. CONCLUSION: The study underlines that work interruptions need to be considered in a much more differentiated way than is currently the case. Both in research and in terms of intervention measures in the work context, the various influencing factors need to be identified for an assessment of the effects on the working person to be possible.

Comparing risk assessment methods for work-related musculoskeletal disorders with in vivo joint loads during manual materials handling.

The validity of observational methods in ergonomics is still challenging research. Criterion validity in terms of concurrent validity is the most commonly studied. However, studies comparing observational methods with biomechanical values are rare. Thus, the aim of this study is to compare the Ovako Working Posture Analysing System (OWAS) and the Rapid Entire Body Assessment (REBA) with in vivo load measurements at hip, spine, and knee during stoop and squat lifting of 14 participants. The results reveal that OWAS and REBA action levels (AL) can distinguish between different in vivo load measurements during manual lifting. However, the results also reveal that the same OWAS- and REBA-AL do not necessarily provide equal mean values of in vivo load measurements. For example, resultant contact force in the vertebral body replacement for squat lifting ranged from 57% body weight (%BW) in OWAS-AL1 to 138%BW in OWAS-AL3 compared to 46%BW in REBA-AL0 and 173%BW in REBA-AL3. Furthermore, the results suggest that the performed squat lifting techniques had a higher risk for work-related musculoskeletal disorders than the performed stoop lifting techniques.

Evolutionary diversity of the control of the azole response by Tra1 across yeast species.

Tra1 is an essential coactivator protein of the yeast SAGA and NuA4 acetyltransferase complexes that regulate gene expression through multiple mechanisms including the acetylation of histone proteins. Tra1 is a pseudokinase of the PIKK family characterized by a C-terminal PI3K domain with no known kinase activity. However, mutations of specific arginine residues to glutamine in the PI3K domains (an allele termed tra1Q3) result in reduced growth and increased sensitivity to multiple stresses. In the opportunistic fungal pathogen Candida albicans, the tra1Q3 allele reduces pathogenicity and increases sensitivity to the echinocandin antifungal drug caspofungin, which disrupts the fungal cell wall. Here, we found that compromised Tra1 function, in contrast to what is seen with caspofungin, increases tolerance to the azole class of antifungal drugs, which inhibits ergosterol synthesis. In C. albicans, tra1Q3 increases the expression of genes linked to azole resistance, such as ERG11 and CDR1. CDR1 encodes a multidrug ABC transporter associated with efflux of multiple xenobiotics, including azoles. Consequently, cells carrying tra1Q3 show reduced intracellular accumulation of fluconazole. In contrast, a tra1Q3 Saccharomyces cerevisiae strain displayed opposite phenotypes: decreased tolerance to azole, decreased expression of the efflux pump PDR5, and increased intracellular accumulation of fluconazole. Therefore, our data provide evidence that Tra1 differentially regulates the antifungal response across yeast species.