mPies: a novel metaproteomics tool for the creation of relevant protein databases and automatized protein annotation

Metaproteomics allows to decipher the structure and functionality of microbial communities. Despite its rapid development, crucial steps such as the creation of standardized protein search databases and reliable protein annotation remain challenging. To overcome those critical steps, we developed a new program named mPies (metaProteomics in environmental sciences). mPies allows the creation of protein databases derived from assembled or unassembled metagenomes, and/or public repositories based on taxon IDs, gene or protein names. For the first time, mPies facilitates the automatization of reliable taxonomic and functional consensus annotations at the protein group level, minimizing the well-known protein inference issue, which is commonly encountered in metaproteomics. mPies’ workflow is highly customizable with regards to input data, workflow steps, and parameter adjustment. mPies is implemented in Python 3/Snakemake and freely available on GitHub: https://github.com/johanneswerner/mPies/

Deciphering the Functioning of Microbial Communities: Shedding Light on the Critical Steps in Metaproteomics

Unraveling the complex structure and functioning of microbial communities is essential to accurately predict the impact of perturbations and/or environmental changes. From all molecular tools available today to resolve the dynamics of microbial communities, metaproteomics stands out, allowing the establishment of phenotype–genotype linkages. Despite its rapid development, this technology has faced many technical challenges that still hamper its potential power. How to maximize the number of protein identification, improve quality of protein annotation, and provide reliable ecological interpretation are questions of immediate urgency. In our study, we used a robust metaproteomic workflow combining two protein fractionation approaches (gel-based versus gel-free) and four protein search databases derived from the same metagenome to analyze the same seawater sample. The resulting eight metaproteomes provided different outcomes in terms of (i) total protein numbers, (ii) taxonomic structures, and (iii) protein functions. The characterization and/or representativeness of numerous proteins from ecologically relevant taxa such as Pelagibacterales, Rhodobacterales, and Synechococcales, as well as crucial environmental processes, such as nutrient uptake, nitrogen assimilation, light harvesting, and oxidative stress response, were found to be particularly affected by the methodology. Our results provide clear evidences that the use of different protein search databases significantly alters the biological conclusions in both gel-free and gel-based approaches. Our findings emphasize the importance of diversifying the experimental workflow for a comprehensive metaproteomic study

Maßnahmen gegen Jakobskreuzkraut Senecio jacobaea - langfristige Aspekte auf Wirksamkeit und biologische Vielfalt

Jakobskreuzkraut (Senecio jacobaea) bereitet zunehmend Probleme auf extensiv geführtem Grünland. Im Jahre 2011 wurde im Rahmen einer Masterarbeit ein Grünland-Großparzellen-Versuch unter Praxisbedingungen angelegt, um die Wirksamkeit verschiedener agronomischer und chemischer Bekämpfungsvarianten gegenüber Senecio jacobaea zu vergleichen. Neben Umbruch mit und ohne Glyphosatbehandlung, wurden Glyphosat mit Neueinsaat (ohne Umbruch) und Simplex (Fluroxypyr+Aminopyralid) mit und ohne Nachsaat geprüft. Ab Herbst 2011 bis 2017 wurden zusätzliche Düngungsvarianten (2 x 50 kg/ha bzw. 1 x 50 kg/ha Kalkammonsalpeter mit 27 % N pro Jahr) in Form von Kleinparzellen in die bestehende Versuchsfläche eingefügt. Seit 2011 wird jährlich die Wirkung der verschiedenen Maßnahmen gegen S. jacobaea bonitiert. Darüber hinaus wird seit 2013 durch Vegetationsaufnahme nach Braun-Blanquet 1964 die Bestandszusammensetzung erhoben. Alle Maßnahmen hatten bis einschließlich 2016 eine dauerhafte Wirkung gegen Jakobskreuzkraut. Darüber hinaus war eine statistisch signifikant verringerte Anzahl an dikotylen Arten feststellbar. Der Unterschied zwischen den Varianten blieb diesbezüglich vergleichsweise gering. Die regelmäßige Düngung führte zu ähnlichen Effekten wie eine einmalige Anwendung von Herbiziden oder Umbruchmaßnahmen. Die stärkste Verringerung der Artenzahl war bei einmaliger Simplex-Anwendung mit nachfolgender regelmäßiger Düngung feststellbar. Die Untersuchungen belegen, dass Jakobskreuzkraut sowohl mit Herbiziden, als auch mit Umbruch oder Düngungsmaßnahmen wirksam zu kontrollieren ist. Allerdings ist dies nicht ohne einen Verlust an Biodiversität (Reduzierung der Artenzahl dikotyler Pflanzen) zu erreichen

Towards the discovery of novel molecular clocks in Prokaryotes

Diel cycle is of enormous biological importance as it imposes daily oscillation in environmental conditions, which temporally structures most ecosystems. Organisms developed biological time-keeping mechanisms – circadian clocks – that provide a significant fitness advantage over competitors by optimising the synchronisation of their biological activities. while circadian clocks are ubiquitous in eukaryotes, they are so far only characterised in Cyanobacteria within Prokaryotes. However, growing evidence suggests that circadian clocks are widespread in the bacterial and archaeal domains. As Prokaryotes are at the heart of crucial environmental processes and are essential to human health, unravelling their time-keeping systems provides numerous applications in medical research, environmental sciences, and biotechnology. in this review, we elaborate on how novel circadian clocks in Prokaryotes offer research and development perspectives. we compare and contrast the different circadian systems in Cyanobacteria and discuss about their evolution and taxonomic distribution. we necessarily provide an updated phylogenetic analysis of bacterial and archaeal species that harbour homologs of the main cyanobacterial clock components. Finally, we elaborate on new potential clock-controlled microorganisms that represent opportunities of ecological and industrial relevance in prokaryotic groups such as anoxygenic photosynthetic bacteria, methanogenic archaea, methanotrophs or sulphate-reducing bacteria

Diel Protein Regulation of Marine Picoplanktonic Communities Assessed by Metaproteomics

The diel cycle is of enormous biological importance in that it imposes temporal structure on ecosystem productivity. In the world’s oceans, microorganisms form complex communities that carry out about half of photosynthesis and the bulk of life-sustaining nutrient cycling. How the functioning of microbial communities is impacted by day and night periods in surface seawater remains to be elucidated. In this study, we compared the day and night metaproteomes of the free-living and the particle-attached bacterial fractions from picoplanktonic communities sampled from the northwest Mediterranean Sea surface. Our results showed similar taxonomic distribution of free-living and particle-attached bacterial populations, with Alphaproteobacteria, Gammaproteobacteria and Cyanobacteria being the most active members. Comparison of the day and night metaproteomes revealed that free-living and particle-attached bacteria were more active during the day and the night, respectively. Interestingly, protein diel variations were observed in the photoautotroph Synechococcales and in (photo)-heterotrophic bacteria such as Flavobacteriales, Pelagibacterales and Rhodobacterales. Moreover, our data demonstrated that diel cycle impacts light-dependent processes such as photosynthesis and UV-stress response in Synechococcales and Rhodobacterales, respectively, while the protein regulation from the ubiquitous Pelagibacterales remained stable over time. This study unravels, for the first time, the diel variation in the protein expression of major free-living and particle-attached microbial players at the sea surface, totaling an analysis of eight metaproteomes

Tensor Graphical Lasso (TeraLasso)

This paper introduces a multi-way tensor generalization of the Bigraphical Lasso (BiGLasso), which uses a two-way sparse Kronecker-sum multivariate-normal model for the precision matrix to parsimoniously model conditional dependence relationships of matrix-variate data based on the Cartesian product of graphs. We call this generalization the {\bf Te}nsor g{\bf ra}phical Lasso (TeraLasso). We demonstrate using theory and examples that the TeraLasso model can be accurately and scalably estimated from very limited data samples of high dimensional variables with multiway coordinates such as space, time and replicates. Statistical consistency and statistical rates of convergence are established for both the BiGLasso and TeraLasso estimators of the precision matrix and estimators of its support (non-sparsity) set, respectively. We propose a scalable composite gradient descent algorithm and analyze the computational convergence rate, showing that the composite gradient descent algorithm is guaranteed to converge at a geometric rate to the global minimizer of the TeraLasso objective function. Finally, we illustrate the TeraLasso using both simulation and experimental data from a meteorological dataset, showing that we can accurately estimate precision matrices and recover meaningful conditional dependency graphs from high dimensional complex datasets.Comment: accepted to JRSS-

Deletion of endogenous Tau proteins is not detrimental in Drosophila

Human Tau (hTau) is a highly soluble and natively unfolded protein that binds to microtubules within neurons. Its dysfunction and aggregation into insoluble paired helical filaments is involved in the pathogenesis of Alzheimer’s disease (AD), constituting, together with accumulated β-amyloid (Aβ) peptides, a hallmark of the disease. Deciphering both the loss-of-function and toxic gain-of-function of hTau proteins is crucial to further understand the mechanisms leading to neurodegeneration in AD. As the fruit fly Drosophila melanogaster expresses Tau proteins (dTau) that are homologous to hTau, we aimed to better comprehend dTau functions by generating a specific tau knock-out (KO) fly line using homologous recombination. We observed that the specific removal of endogenous dTau proteins did not lead to overt, macroscopic phenotypes in flies. Indeed, survival, climbing ability and neuronal function were unchanged in tau KO flies. In addition, we did not find any overt positive or negative effect of dTau removal on human Aβ-induced toxicity. Altogether, our results indicate that the absence of dTau proteins has no major functional impact on flies, and suggests that our tau KO strain is a relevant model to further investigate the role of dTau proteins in vivo, thereby giving additional insights into hTau functions