Evaluated articles

Evaluation Summary:

Since the inception of comparative genomics, mining phyletic patterns has been a powerful approach for the discovery of previously unknown biological interactions. The authors use a combination of singular value decomposition of the phyletic pattern matrix and random forests classification method to uncover potential protein-protein interactions. The work illustrates the utility of such methods, which are finding increasing application in addressing various computational biological problems, such as predicting protein-protein interactions from genomic information.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. The reviewers remained anonymous to the authors.)

Evaluation Summary:

Toxoplasma gondii is a widespread parasite of warm blooded animals, with estimates suggesting 2 billion people are currently and chronically infected with this pathogen. Many questions remain as to how humans control and eliminate T. gondii following infection. In this manuscript, Rinkenberger et al. reveal a previously unidentified and understudied host factor, RARRES3 that promotes cell autonomous control of T. gondii in human cells. The precise mechanism of control and its in vivo relevance remain areas for additional work.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #2 agreed to share their name with the authors.)

Evaluation Summary:

This study will interest neuroscientists, in particular those interested in the neurocircuitry of emotional behaviors. Using modern neuroscience techniques, the authors demonstrate that anatomical projections from a brain stem structure called the parabrachial nucleus to the paraventricular nucleus thalamus contribute to aversive states like fear and anxiety. Overall, the study offers important details of a previously uncharacterized brain circuit, although some additional experiments are required to fully substantiate the authors' claims.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #1 and Reviewer #2 agreed to share their names with the authors.)

Evaluation Summary:

The paper by Rai and colleagues examines the transcriptional response of Candida glabrata, a common human fungal pathogen, during interaction with macrophages. They use RNA PolII profiling to identify not just the total transcripts but instead focus on the actively transcribing genes. By examining the profile over time, they identify particular transcripts that are enriched at each time point, building a hierarchical model for how a transcription factor, CgXbp1, may regulate part of this response. While the authors have generated a large and potentially impactful dataset, along with several interesting observations, it is important to be cautious as the direct targets of CgXbp1 were characterized under one particular condition and the transcriptional analyses were obtained in another condition, one shown to be highly dynamic as during macrophage infection.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. The reviewers remained anonymous to the authors.)

Evaluation Summary:

The paper describes single-molecule experiments that address the assembly of a double hexamer of the Mcm2-7 complex that is required to license all origins of DNA replication in eukaryotic cells by formation of a pre-Replicative Complex (pre-RC). The observations show that one Origin Recognition Complex, an ATP-dependent DNA binding protein, can load both Mcm2-7 hexamers in opposite orientation. The results nicely complement prior data on the mechanism of pre-RC assembly.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #1 agreed to share their name with the authors.)

Evaluation Summary:

This manuscript will be of interest for scientists interested in cell cycle, DNA repair, and genome stability reporting the unexpected discovery that the DNA-dependent protein kinase (DNA-PK) is required for DSB resection in G0 cells, whereas it is known and confirmed here that it inhibits resection in G1 and G2 cells. This finding has important implications for the clinical application of DNA-PK-targeted inhibitors. The data are of high quality and derive from two independent cell lines, genetic requirements were mostly established by gene knockouts, and the latest genome-wide sequencing techniques were applied to measure resection tracts. The key claims of the manuscript are supported by the data presented by the authors; however, further validations are needed to strengthen the quality and impact of the paper.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #1 agreed to share their name with the authors.)

Evaluation Summary:

This study fuses images from cardiac magnetic resonance imaging and T1-mapping to reconstruct 3D anatomical models of the heart from hypertrophic cardiomyopathy patients. Using the model, they investigated potential contributions of diffuse fibrosis to arrhythmogenesis of the heart model in response to focal stimulation. While not perfect, the computer model significantly outperforms other risk predictors, and highlights diffuse fibrosis as a possible underlying cause. This study will be of interest to clinicians and basic scientists involved in heart rhythm research.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #3 agreed to share their name with the authors.)

Evaluation Summary:

This manuscript investigates the cellular role of the juxtamembrane region in the EGF receptor, a poorly understood portion of the EGFR cytosolic domain that connects the transmembrane segment to the kinase domain. Through a series of well-designed experiments, the work shows that the endocytic trafficking route of EGFR following its activation is determined by the juxtamembrane coiled-coil conformation in a model cell line. This finding is important for three reasons. It identifies a critical role for the juxtamembrane region; it resolves the discrepancy that TGF-beta dissociation from EGFR is supposed to occur at higher pH, yet the EGFR-TGF-beta complex continues to signal from endosomes; and it pinpoints the mechanism of EGFR inhibition by a new class of tyrosine kinase inhibitors.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. All reviewers agreed to share their names with the authors.)

Evaluation Summary:

Kunze et al. provide a fine experiment to show that both increases in mean temperature and (extreme) variability in temperature regimes have important consequences in host-pathogen interactions. The results presented in this manuscript shed a light on why disease spread models fed by experimental data (commonly obtained in stable environmental conditions) are frequently inaccurate. These results lead us to more realistic understanding of the impacts of climate change in biological species but also identify the need of mechanisms behind species interaction in fluctuating environments/temperatures. This manuscript thus comes timely as the planet is warming, and disease ecologists, limnologists, epidemiologists and physiologists are interested in the consequences.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. The reviewers remained anonymous to the authors).

Evaluation Summary:

This paper represents multiple milestones in our understanding of the evolution and extinction of Pleistocene equids, including revising the timing of extinction and clarifying the evolutionary history of Equus (Sussemionus) ovodovi. The discovery of the late persistence of non-caballine equid taxa in northern China until deep into the late Holocene is particularly important. This finding will be of broad interest to the paleontology, paleoecology, archaeology, paleogenomic communities and should stimulate important future research into equid extinction processes.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. The reviewers remained anonymous to the authors.)

Evaluation Summary:

Multiple inherited mutations in the epithelial CFTR anion-permeable channel cause cystic fibrosis through different molecular mechanisms that can be targeted by different types of drugs to treat the disease. Drawing from available structural information and double-mutant cycle analysis of patch-clamp recordings, Simon and Csanády find that one of the most common CFTR disease-causing mutations, R117H, disrupts an interaction between the R117 side-chain and a main-chain carbonyl that selectively stabilizes the open state of the channel. These findings may open new paths of exploration for treating patients carrying this mutation, and provide important mechanistic constraints towards understanding the gating mechanism of CFTR proteins.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #1 and Reviewer #2 agreed to share their names with the authors.)

Evaluation Summary:

Homeostatic plasticity helps to confine neural network activity within limits. In this study, the authors show that loss of PAR bZIP family of transcription factors leads to overcompensation of excitatory synaptic transmission and average network activity upon sustained activity deprivation. The work identifies an endogenous transcriptional program that constrains upward homeostatic response and whose activity is implicated in preventing aberrant network activity associated with epilepsy and other brain disorders. These are exciting results that address the question of broad importance. While most arguments are supported by data of high quality, further experiments would strengthen the claims about the relative contribution of excitatory and inhibitory mechanisms and clarify the nature of compensation.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #1 agreed to share their name with the authors.)

Evaluation Summary:

This paper will be of broad interest to many fields, including drug discovery, cancer biology, and structure biology. It makes a significant advance in understanding the mechanism of action of hormone therapies for breast cancer, and how resistance driving mutations alter drug responses. The structural biology data has clear potential for strong impact though some additional analysis might be needed.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #2 agreed to share their name with the authors.)

Evaluation Summary:

This manuscript focusses on a little studied, but highly interesting presumptive mechanosensory cell type in cnidarians, the 'hair cell'. The work shows that the POU-IV transcription factor is required for the maturation of this cell type in the sea anemone Nematostella vectensis. Because POU-IV transcription factors also play essential roles in the differentiation of mechanoreceptors in many bilaterian phyla, this suggests an evolutionarily ancient role of POU-IV in regulating mechanosensory identity. This study will hence be of great interest to developmental biologists and evolutionary biologists who are interested in the developmental evolution of neuronal cell types.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #1, Reviewer #2 and Reviewer #3 agreed to share their name with the authors.)

Evaluation Summary:

Cholesterol has long been known to have significant effects on G protein-coupled receptor (GPCR) ligand binding properties and stability, and cholesterol/GPCR interactions have frequently been observed in high-resolution structures. However, relatively limited biophysical work has been done to investigate the mechanistic basis for cholesterol's effects. This manuscript describes the use of a sensitive 19F NMR probe to monitor conformational equilibria in a prototypical GPCR, the A2a adenosine receptor. These experiments, together with data from other NMR experiments, computational analysis, and G protein assays, show that the subtle effects of cholesterol derive in large part from modulation of membrane biophysical properties, in contrast to conventional allosteric modulators that exert their effects through direct long-lived receptor binding.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #2 agreed to share their name with the authors.)

Evaluation Summary:

Smith et al. describes the propagation patterns of electrical activity in the brains of human epileptic patients. The authors demonstrate that interictal spikes, commonly observed electrical events in epileptic patients, propagate in a similar manner to seizures. This suggests that interictal spikes could be used in surgical planning, which would be of great interest to neurosurgeons and neurologists treating patients with medication refractory epilepsy.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #1 agreed to share their name with the authors.)

Evaluation Summary:

This manuscript is of broad interest to developmental biologists and those studying transcriptional/epigenetic regulation of cell-specific and housekeeping gene programs. The work demonstrates that Polycomb complexes coordinate the regulation of distinct groups of genes during early embryogenesis, which offers interesting insights into how very early embryos differentially control housekeeping versus specific developmental gene promoters/enhancers. The data are of high quality, and the conclusions are insightful yet measured.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #3 agreed to share their name with the authors.)

Evaluation Summary:

Kröger et al use 2-photon FLIM tomography to perform correlative imaging on in vitro, ex vivo and in vivo blood and skin cells to determine characteristic NADPH fluorescence lifetimes for M1 and M2 ends of macrophage spectrum. Interestingly, M1 and M2 macrophages, and all other tissue cells, had distinctive lifetime features leading to robust prediction of phenotypes, with ground trust defined by cytokine staining. They generate a decision tree that has ~90% accuracy in identifying M1 and M2 based on FLIM parameters and additional information. The ability to use two photon fluorescence lifetime tomography of NADPH fluorescence to identify macrophages and their inflammatory status in human tissues should open opportunities in experimental medicine and eventually medical diagnosis.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #1 agreed to share their name with the authors.)

Evaluation Summary:

This paper is of outstanding interest to the broad community of scientists interested in biological energy conversion in general and rotary ATPases in particular. The authors show that the 36{degree sign} power stroke in ATP synthesis is subdivided into two steps of 11{degree sign} and 25{degree sign} in the E. coli enzyme, which serves as a comparatively simple model of the fundamental and universally important process of ATP production in mitochondria and chloroplasts. By combining precise and sophisticated single-molecule studies with directed mutagenesis, this work provides the much-needed functional context for recent high-resolution cryo-EM structures of rotary ATPases.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #2 agreed to share their name with the authors.)

Evaluation Summary:

This paper is of interest not only for immunologists studying the inflammation, but also for biomedical researchers studying various biological processes using C57BL/6 mice. The data in this paper indicate that genetic differences between C57BL/6 substrains can affect reproducibility and generalizability in a broad range of biological studies with mouse models reported to date.

(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. The reviewers remained anonymous to the authors.)