Cells use homology-dependent DNA repair to mend chromosome breaks and restore broken replication forks, thereby ensuring genome stability and cell survival. DNA break repair via homology-based mechanisms involves nuclease-dependent DNA end resection, which generates long tracts of single-stranded DNA required for checkpoint activation and loading of homologous recombination proteins Rad52/51/55/57. While recruitment of the homologous recombination machinery is well characterized, it is not known how its presence at repair loci is coordinated with downstream re-synthesis of resected DNA We show that Rad51 inhibits recruitment of proliferating cell nuclear antigen (PCNA), the platform for assembly of the DNA replication machinery, and that unloading of Rad51 by Srs2 helicase is required for efficient PCNA loading and restoration of resected DNA As a result, srs2Δ mutants are deficient in DNA repair correlating with extensive DNA processing, but this defect in srs2Δ mutants can be suppressed by inactivation of the resection nuclease Exo1. We propose a model in which during re-synthesis of resected DNA, the replication machinery must catch up with the preceding processing nucleases, in order to close the single-stranded gap and terminate further resection.

• MeSH
• biologické modely MeSH
• DNA metabolismus   MeSH
• enzymy opravy DNA metabolismus   MeSH
• homologní rekombinace * MeSH
• poškození DNA * MeSH
• proliferační antigen buněčného jádra metabolismus   MeSH
• rekombinační oprava DNA * MeSH
• rekombinasy metabolismus   MeSH
• Saccharomyces cerevisiae enzymologie   genetika   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH

Protein modifications regulate both DNA repair levels and pathway choice. How each modification achieves regulatory effects and how different modifications collaborate with each other are important questions to be answered. Here, we show that sumoylation regulates double-strand break repair partly by modifying the end resection factor Sae2. This modification is conserved from yeast to humans, and is induced by DNA damage. We mapped the sumoylation site of Sae2 to a single lysine in its self-association domain. Abolishing Sae2 sumoylation by mutating this lysine to arginine impaired Sae2 function in the processing and repair of multiple types of DNA breaks. We found that Sae2 sumoylation occurs independently of its phosphorylation, and the two modifications act in synergy to increase soluble forms of Sae2. We also provide evidence that sumoylation of the Sae2-binding nuclease, the Mre11-Rad50-Xrs2 complex, further increases end resection. These findings reveal a novel role for sumoylation in DNA repair by regulating the solubility of an end resection factor. They also show that collaboration between different modifications and among multiple substrates leads to a stronger biological effect.

• MeSH
• DNA vazebné proteiny genetika   MeSH
• dvouřetězcové zlomy DNA MeSH
• endodeoxyribonukleasy genetika   MeSH
• endonukleasy genetika   MeSH
• exodeoxyribonukleasy genetika   MeSH
• fosforylace MeSH
• lidé MeSH
• oprava DNA spojením konců genetika   MeSH
• oprava DNA genetika   MeSH
• poškození DNA genetika   MeSH
• rozpustnost MeSH
• Saccharomyces cerevisiae - proteiny genetika   MeSH
• Saccharomyces cerevisiae MeSH
• sumoylace genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

Membrane-bound pyrophosphatases couple the hydrolysis of inorganic pyrophosphate to the pumping of ions (sodium or protons) across a membrane in order to generate an electrochemical gradient. This class of membrane protein is widely conserved across plants, fungi, archaea, and bacteria, but absent in multicellular animals, making them a viable target for drug design against protozoan parasites such as Plasmodium falciparum. An excellent understanding of many of the catalytic states throughout the enzymatic cycle has already been afforded by crystallography. However, the dynamics and kinetics of the catalytic cycle between these static snapshots remain to be elucidated. Here, we employ single-molecule Förster resonance energy transfer (FRET) measurements to determine the dynamic range and frequency of conformations available to the enzyme in a lipid bilayer during the catalytic cycle. First, we explore issues related to the introduction of fluorescent dyes by cysteine mutagenesis; we discuss the importance of residue selection for dye attachment, and the balance between mutating areas of the protein that will provide useful dynamics while not altering highly conserved residues that could disrupt protein function. To complement and guide the experiments, we used all-atom molecular dynamics simulations and computational methods to estimate FRET efficiency distributions for dye pairs at different sites in different protein conformational states. We present preliminary single-molecule FRET data that points to insights about the binding modes of different membrane-bound pyrophosphatase substrates and inhibitors.

• MeSH
• bakteriální proteiny chemie   genetika   izolace a purifikace   metabolismus   MeSH
• buněčná membrána metabolismus   MeSH
• enzymatické testy přístrojové vybavení   metody   MeSH
• fluorescenční barviva chemie   MeSH
• fluorescenční mikroskopie přístrojové vybavení   metody   MeSH
• mutageneze MeSH
• protozoální proteiny chemie   genetika   izolace a purifikace   metabolismus   MeSH
• pyrofosfatasy chemie   genetika   izolace a purifikace   metabolismus   MeSH
• racionální návrh léčiv MeSH
• rekombinantní proteiny chemie   genetika   izolace a purifikace   metabolismus   MeSH
• rezonanční přenos fluorescenční energie přístrojové vybavení   metody   MeSH
• Saccharomyces cerevisiae MeSH
• sekvenční seřazení MeSH
• simulace molekulární dynamiky * MeSH
• software MeSH
• zobrazení jednotlivé molekuly přístrojové vybavení   metody   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In yeast, phosphatidylglycerol (PG) is a minor phospholipid under standard conditions; it can be utilized for cardiolipin (CL) biosynthesis by CL synthase, Crd1p, or alternatively degraded by the phospholipase Pgc1p. The Saccharomyces cerevisiae deletion mutants crd1Δ and pgc1Δ both accumulate PG. Based on analyses of the phospholipid content of pgc1Δ and crd1Δ yeast, we revealed that in yeast mitochondria, two separate pools of PG are present, which differ in their fatty acid composition and accessibility for Pgc1p-catalyzed degradation. In contrast to CL-deficient crd1Δ yeast, the pgc1Δ mutant contains normal levels of CL. This makes the pgc1Δ strain a suitable model to study the effect of accumulation of PG per se. Using fluorescence microscopy, we show that accumulation of PG with normal levels of CL resulted in increased fragmentation of mitochondria, while in the absence of CL, accumulation of PG led to the formation of large mitochondrial sheets. We also show that pgc1Δ mitochondria exhibited increased respiration rates due to increased activity of cytochrome c oxidase. Taken together, our results indicate that not only a lack of anionic phospholipids, but also excess PG, or unbalanced ratios of anionic phospholipids in mitochondrial membranes, have harmful consequences on mitochondrial morphology and function.

• MeSH
• fosfatidylglyceroly metabolismus   MeSH
• fosfolipasy fyziologie   MeSH
• kardiolipiny biosyntéza   MeSH
• mitochondrie metabolismus   ultrastruktura   MeSH
• respirační komplex IV metabolismus   MeSH
• Saccharomyces cerevisiae metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

As species recover from exploitation, continued assessments of connectivity and population structure are warranted to provide information for conservation and management. This is particularly true in species with high dispersal capacity, such as migratory whales, where patterns of connectivity could change rapidly. Here we build on a previous long-term, large-scale collaboration on southern right whales (Eubalaena australis) to combine new (nnew) and published (npub) mitochondrial (mtDNA) and microsatellite genetic data from all major wintering grounds and, uniquely, the South Georgia (Islas Georgias del Sur: SG) feeding grounds. Specifically, we include data from Argentina (npub mtDNA/microsatellite = 208/46), Brazil (nnew mtDNA/microsatellite = 50/50), South Africa (nnew mtDNA/microsatellite = 66/77, npub mtDNA/microsatellite = 350/47), Chile-Peru (nnew mtDNA/microsatellite = 1/1), the Indo-Pacific (npub mtDNA/microsatellite = 769/126), and SG (npub mtDNA/microsatellite = 8/0, nnew mtDNA/microsatellite = 3/11) to investigate the position of previously unstudied habitats in the migratory network: Brazil, SG, and Chile-Peru. These new genetic data show connectivity between Brazil and Argentina, exemplified by weak genetic differentiation and the movement of 1 genetically identified individual between the South American grounds. The single sample from Chile-Peru had an mtDNA haplotype previously only observed in the Indo-Pacific and had a nuclear genotype that appeared admixed between the Indo-Pacific and South Atlantic, based on genetic clustering and assignment algorithms. The SG samples were clearly South Atlantic and were more similar to the South American than the South African wintering grounds. This study highlights how international collaborations are critical to provide context for emerging or recovering regions, like the SG feeding ground, as well as those that remain critically endangered, such as Chile-Peru.

• MeSH
• genetická variace * MeSH
• genotypizační techniky MeSH
• migrace zvířat MeSH
• ostrovy MeSH
• rozšíření zvířat MeSH
• stravovací zvyklosti MeSH
• velryby genetika   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Brazílie MeSH
• Chile MeSH
• ostrovy MeSH
• Peru MeSH

A must-have resource for any orthopaedic library, the latest edition of this technique-focused guide to the elbow has been revised and updated to give you even more coverage of trauma, arthroscopy, soft tissue injury, and joint replacement. The new full-color illustrations and online access to 43 video clips of exams and procedures performed by experts visually enhance an already great resource for both the novice becoming familiar with elbow anatomy and biomechanics and the seasoned surgeon treating difficult elbow problems.

• MeSH
• artroplastiky kloubů MeSH
• artroskopie MeSH
• loketní kloub chirurgie   MeSH
• nemoci kloubů chirurgie   terapie   MeSH
• poranění lokte MeSH
• totální endoprotéza loketního kloubu MeSH
• záchranná terapie MeSH

• Konspekt
• Ortopedie. Chirurgie. Oftalmologie
• NLK Obory
• ortopedie
• NLK Publikační typ
• kolektivní monografie

Collected papers from a symposium that focused on endothelium-dependent hyperpolarization. Intended for professional public.

• MeSH
• cévní endotel cytologie   účinky léků   MeSH
• farmakoterapie MeSH
• krevní oběh účinky léků   MeSH
• krevní tlak účinky léků   MeSH
• svaly hladké cévní cytologie   účinky léků   MeSH
• vazodilatace MeSH
• vazodilatancia terapeutické užití   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• kongresy MeSH
• sborníky MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• angiologie
• farmacie a farmakologie

In recent years, the scientific community has called for improvements in the credibility, robustness and reproducibility of research, characterized by increased interest and promotion of open and transparent research practices. While progress has been positive, there is a lack of consideration about how this approach can be embedded into undergraduate and postgraduate research training. Specifically, a critical overview of the literature which investigates how integrating open and reproducible science may influence student outcomes is needed. In this paper, we provide the first critical review of literature surrounding the integration of open and reproducible scholarship into teaching and learning and its associated outcomes in students. Our review highlighted how embedding open and reproducible scholarship appears to be associated with (i) students' scientific literacies (i.e. students' understanding of open research, consumption of science and the development of transferable skills); (ii) student engagement (i.e. motivation and engagement with learning, collaboration and engagement in open research) and (iii) students' attitudes towards science (i.e. trust in science and confidence in research findings). However, our review also identified a need for more robust and rigorous methods within pedagogical research, including more interventional and experimental evaluations of teaching practice. We discuss implications for teaching and learning scholarship.

• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Plant traits-the morphological, anatomical, physiological, biochemical and phenological characteristics of plants-determine how plants respond to environmental factors, affect other trophic levels, and influence ecosystem properties and their benefits and detriments to people. Plant trait data thus represent the basis for a vast area of research spanning from evolutionary biology, community and functional ecology, to biodiversity conservation, ecosystem and landscape management, restoration, biogeography and earth system modelling. Since its foundation in 2007, the TRY database of plant traits has grown continuously. It now provides unprecedented data coverage under an open access data policy and is the main plant trait database used by the research community worldwide. Increasingly, the TRY database also supports new frontiers of trait-based plant research, including the identification of data gaps and the subsequent mobilization or measurement of new data. To support this development, in this article we evaluate the extent of the trait data compiled in TRY and analyse emerging patterns of data coverage and representativeness. Best species coverage is achieved for categorical traits-almost complete coverage for 'plant growth form'. However, most traits relevant for ecology and vegetation modelling are characterized by continuous intraspecific variation and trait-environmental relationships. These traits have to be measured on individual plants in their respective environment. Despite unprecedented data coverage, we observe a humbling lack of completeness and representativeness of these continuous traits in many aspects. We, therefore, conclude that reducing data gaps and biases in the TRY database remains a key challenge and requires a coordinated approach to data mobilization and trait measurements. This can only be achieved in collaboration with other initiatives.

• MeSH
• biodiverzita MeSH
• ekologie MeSH
• ekosystém * MeSH
• přístup k informacím * MeSH
• rostliny MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• bolest rehabilitace   MeSH
• management bolesti MeSH
• muskuloskeletální nemoci rehabilitace   MeSH

• Konspekt
• Ortopedie. Chirurgie. Oftalmologie
• NLK Obory
• ortopedie
• rehabilitační a fyzikální medicína
• NLK Publikační typ
• kolektivní monografie