Data‐driven modeling reconciles kinetics of ERK phosphorylation,localization, and activity states

The extracellular signal‐regulated kinase (ERK) signaling pathway controls cell proliferation and differentiation in metazoans. Two hallmarks of its dynamics are adaptation of ERK phosphorylation, which has been linked to negative feedback, and nucleocytoplasmic shuttling, which allows active ERK to phosphorylate protein substrates in the nucleus and cytosol. To integrate these complex features, we acquired quantitative biochemical and live‐cell microscopy data to reconcile phosphorylation, localization, and activity states of ERK. While maximal growth factor stimulation elicits transient ERK phosphorylation and nuclear translocation responses, ERK activities available to phosphorylate substrates in the cytosol and nuclei show relatively little or no adaptation. Free ERK activity in the nucleus temporally lags the peak in nuclear translocation, indicating a slow process. Additional experiments, guided by kinetic modeling, show that this process is consistent with ERK's modification of and release from nuclear substrate anchors. Thus, adaptation of whole‐cell ERK phosphorylation is a by‐product of transient protection from phosphatases. Consistent with this interpretation, predictions concerning the dose‐dependence of the pathway response and its interruption by inhibition of MEK were experimentally confirmed.     

Dynameomics: Data‐driven methods and models for utilizing large‐scale protein structure repositories for improving fragment‐based loop prediction

Protein function is intimately linked to protein structure and dynamics yet experimentally determined structures frequently omit regions within a protein due to indeterminate data, which is often due protein dynamics. We propose that atomistic molecular dynamics simulations provide a diverse sampling of biologically relevant structures for these missing segments (and beyond) to improve structural modeling and structure prediction. Here we make use of the Dynameomics data warehouse, which contains simulations of representatives of essentially all known protein folds. We developed novel computational methods to efficiently identify, rank and retrieve small peptide structures, or fragments, from this database. We also created a novel data model to analyze and compare large repositories of structural data, such as contained within the Protein Data Bank and the Dynameomics data warehouse. Our evaluation compares these structural repositories for improving loop predictions and analyzes the utility of our methods and models. Using a standard set of loop structures, containing 510 loops, 30 for each loop length from 4 to 20 residues, we find that the inclusion of Dynameomics structures in fragment‐based methods improves the quality of the loop predictions without being dependent on sequence homology. Depending on loop length, ～25–75% of the best predictions came from the Dynameomics set, resulting in lower main chain root‐mean‐square deviations for all fragment lengths using the combined fragment library. We also provide specific cases where Dynameomics fragments provide better predictions for NMR loop structures than fragments from crystal structures. Online access to these fragment libraries is available at http://www.dynameomics.org/fragments .     

Data‐independent acquisition‐based SWATH‐MS for quantitative proteomics: a tutorial

Many research questions in fields such as personalized medicine, drug screens or systems biology depend on obtaining consistent and quantitatively accurate proteomics data from many samples. SWATH‐MS is a specific variant of data‐independent acquisition (DIA) methods and is emerging as a technology that combines deep proteome coverage capabilities with quantitative consistency and accuracy. In a SWATH‐MS measurement, all ionized peptides of a given sample that fall within a specified mass range are fragmented in a systematic and unbiased fashion using rather large precursor isolation windows. To analyse SWATH‐MS data, a strategy based on peptide‐centric scoring has been established, which typically requires prior knowledge about the chromatographic and mass spectrometric behaviour of peptides of interest in the form of spectral libraries and peptide query parameters. This tutorial provides guidelines on how to set up and plan a SWATH‐MS experiment, how to perform the mass spectrometric measurement and how to analyse SWATH‐MS data using peptide‐centric scoring. Furthermore, concepts on how to improve SWATH‐MS data acquisition, potential trade‐offs of parameter settings and alternative data analysis strategies are discussed.     

Alternative fire‐driven vegetation states

There is increasing evidence that alternative stable vegetation types exist for a given climate that are maintained by distinct fire regimes. Paritsis et al. (2014, this issue) provide an example in a temperate ecosystem. Here I briefly review cases of bi‐stability in various climates, and present a simple model for the transition between states in their system.     

Projected climate‐driven faunal movement routes

Historically, many species moved great distances as climates changed. However, modern movements will be limited by the patterns of human‐dominated landscapes. Here, we use a combination of projected climate‐driven shifts in the distributions of 2903 vertebrate species, estimated current human impacts on the landscape, and movement models, to determine through which areas in the western hemisphere species will likely need to move to track suitable climates. Our results reveal areas with projected high densities of climate‐driven movements – including, the Amazon Basin, the southeastern United States and southeastern Brazil. Some of these regions, such as southern Bolivia and northern Paraguay, contain relatively intact landscapes, whereas others such as the southeastern United States and Brazil are heavily impacted by human activities. Thus, these results highlight both critical areas for protecting lands that will foster movement, and barriers where human land‐use activities will likely impede climate‐driven shifts in species distributions.     

Sirt1 protects from K‐Ras‐driven lung carcinogenesis

The NAD⁺‐dependent deacetylase SIRT1 can be oncogenic or tumor suppressive depending on the tissue. Little is known about the role of SIRT1 in non‐small cell lung carcinoma (NSCLC), one of the deadliest cancers, that is frequently associated with mutated K‐RAS. Therefore, we investigated the effect of SIRT1 on K‐RAS‐driven lung carcinogenesis. We report that SIRT1 protein levels are downregulated by oncogenic K‐RAS in a MEK and PI3K‐dependent manner in mouse embryo fibroblasts (MEFs), and in human lung adenocarcinoma cell lines. Furthermore, Sirt1 overexpression in mice delays the appearance of K‐Ras^G12V‐driven lung adenocarcinomas, reducing the number and size of carcinomas at the time of death and extending survival. Consistently, lower levels of SIRT1 are associated with worse prognosis in human NSCLCs. Mechanistically, analysis of mouse Sirt1‐Tg pneumocytes, isolated shortly after K‐Ras^G12V activation, reveals that Sirt1 overexpression alters pathways involved in tumor development: proliferation, apoptosis, or extracellular matrix organization. Our work demonstrates a tumor suppressive role of SIRT1 in the development of K‐RAS‐driven lung adenocarcinomas in mice and humans, suggesting that the SIRT1–K‐RAS axis could be a therapeutic target for NSCLCs.     

Behaviour‐driven micro‐scale niche differentiation in carabid beetles

Carabid beetles form rich and abundant communities in arable landscapes. Their generalist feeding behaviour and similar environmental requirements raise questions about the mechanisms allowing the coexistence of such species‐rich assemblages. We hypothesized that subtle niche partitioning comes into play on spatial, temporal, or trophic basis. To test this, we performed experiments and made observations on the behaviour of two sympatric carabid species of similar size and life cycle, Bembidion quadrimaculatum L. and Phyla obtusa Audinet‐Serville (both Coleoptera: Carabidae: Bembidiini). We compared plant climbing behaviour, daily activity patterns, and trophic preferences between the two carabid species under laboratory conditions. Whereas no clear difference in trophic preference was observed, our results suggest temporal niche differentiation at the nychthemeron scale (a period of 24 consecutive hours), with one of the species being more diurnal and the other more nocturnal, and spatial differentiation in their habitat use at the plant stratum scale. Intra‐specific variation suggests that micro‐scale spatio‐temporal niche differentiation could be mediated by behavioural plasticity in these two carabid species. We speculate that such behavioural plasticity may provide carabid beetles with a high adaptive potential in intensively managed agricultural areas.     

Exploiting the Link between Protein Rigidity and Thermostability for Data‐Driven Protein Engineering

Understanding and exploiting the relationship between microscopic structure and macroscopic stability is important for developing strategies to improve protein stability at high temperatures. The thermostability of proteins has been repeatedly linked to an enhanced structural rigidity of the folded native state. In the current study, the rigidity of protein structures from mesophilic and thermophilic organisms along a thermal unfolding trajectory is directly probed. In order to perform this, protein structures were modeled as constraint networks, and the rigidity in these networks was quantified using the Floppy Inclusion and Rigid Substructure Topography (FIRST) method. During the thermal unfolding, a phase transition was observed that defines the rigidity percolation threshold and corresponds to the folded‐unfolded transition in protein folding. Using concepts from percolation theory and network science, a higher phase transition temperature was observed for ca. two‐thirds of the proteins from thermophilic organisms compared to their mesophilic counterparts, when applied to a data set of 20 pairs of homologues. From both the analysis of the microstructure of the constraint networks and monitoring the macroscopic behavior during the thermal unfolding, direct evidence was found for the “corresponding states” concept, which states that mesophilic and thermophilic enzymes are in corresponding states of similar flexibility at their respective optimal temperature. Finally, the current approach facilitated the identification of structural features from which a destabilization of the structure originates upon thermal unfolding. These predictions show a good agreement with the experimental data. Therefore, the information might be exploited in data‐driven protein engineering by pointing to residues that should be varied to obtain a protein with higher thermostability.     

A Semiparametric Missing‐Data‐Induced Intensity Method for Missing Covariate Data in Individually Matched Case–Control Studies

Summary In individually matched case–control studies, when some covariates are incomplete, an analysis based on the complete data may result in a large loss of information both in the missing and completely observed variables. This usually results in a bias and loss of efficiency. In this article, we propose a new method for handling the problem of missing covariate data based on a missing‐data‐induced intensity approach when the missingness mechanism does not depend on case–control status and show that this leads to a generalization of the missing indicator method. We derive the asymptotic properties of the estimates from the proposed method and, using an extensive simulation study, assess the finite sample performance in terms of bias, efficiency, and 95% confidence coverage under several missing data scenarios. We also make comparisons with complete‐case analysis (CCA) and some missing data methods that have been proposed previously. Our results indicate that, under the assumption of predictable missingness, the suggested method provides valid estimation of parameters, is more efficient than CCA, and is competitive with other, more complex methods of analysis. A case–control study of multiple myeloma risk and a polymorphism in the receptor Inter‐Leukin‐6 (IL‐6‐α) is used to illustrate our findings.     

ParA‐mediated plasmid partition driven by protein pattern self‐organization

DNA segregation ensures the stable inheritance of genetic material prior to cell division. Many bacterial chromosomes and low‐copy plasmids, such as the plasmids P1 and F, employ a three‐component system to partition replicated genomes: a partition site on the DNA target, typically called parS, a partition site binding protein, typically called ParB, and a Walker‐type ATPase, typically called ParA, which also binds non‐specific DNA. In vivo, the ParA family of ATPases forms dynamic patterns over the nucleoid, but how ATP‐driven patterning is involved in partition is unknown. We reconstituted and visualized ParA‐mediated plasmid partition inside a DNA‐carpeted flowcell, which acts as an artificial nucleoid. ParA and ParB transiently bridged plasmid to the DNA carpet. ParB‐stimulated ATP hydrolysis by ParA resulted in ParA disassembly from the bridging complex and from the surrounding DNA carpet, which led to plasmid detachment. Our results support a diffusion‐ratchet model, where ParB on the plasmid chases and redistributes the ParA gradient on the nucleoid, which in turn mobilizes the plasmid.     

Light‐driven tipping points in polar ecosystems

Some ecosystems can undergo abrupt transformation in response to relatively small environmental change. Identifying imminent ‘tipping points’ is crucial for biodiversity conservation, particularly in the face of climate change. Here, we describe a tipping point mechanism likely to induce widespread regime shifts in polar ecosystems. Seasonal snow and ice‐cover periodically block sunlight reaching polar ecosystems, but the effect of this on annual light depends critically on the timing of cover within the annual solar cycle. At high latitudes, sunlight is strongly seasonal, and ice‐free days around the summer solstice receive orders of magnitude more light than those in winter. Early melt that brings the date of ice‐loss closer to midsummer will cause an exponential increase in the amount of sunlight reaching some ecosystems per year. This is likely to drive ecological tipping points in which primary producers (plants and algae) flourish and out‐compete dark‐adapted communities. We demonstrate this principle on Antarctic shallow seabed ecosystems, which our data suggest are sensitive to small changes in the timing of sea‐ice loss. Algae respond to light thresholds that are easily exceeded by a slight reduction in sea‐ice duration. Earlier sea‐ice loss is likely to cause extensive regime shifts in which endemic shallow‐water invertebrate communities are replaced by algae, reducing coastal biodiversity and fundamentally changing ecosystem functioning. Modeling shows that recent changes in ice and snow cover have already transformed annual light budgets in large areas of the Arctic and Antarctic, and both aquatic and terrestrial ecosystems are likely to experience further significant change in light. The interaction between ice‐loss and solar irradiance renders polar ecosystems acutely vulnerable to abrupt ecosystem change, as light‐driven tipping points are readily breached by relatively slight shifts in the timing of snow and ice‐loss.