Cell Biology

Evaluation Summary:

Over the past decade, the role of ATP levels in the material properties of cells has gathered substantial interest in part because of the potential role of ATP in solubilizing biomolecular condensates. This study uses a quantitative imaging-based measurement of ATP levels in live cells to assess the impact of mutants in ATP homeostasis on ATP levels and protein aggregation. The strength of this paper is the quantitative, single cell analysis, and the manipulation of ATP using native control pathways. The authors suggest that fluctuations in ATP concentrations can lead to protein aggregation, which would be of broad interest to many fields, including cell biology, aging and neurodegeneration.

(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 study focuses on the formation of adaptor protein complexes at adhesion sites and their links to in vitro membrane ruffling and cell movement. Specifically, the authors study the role of the adaptor BCAR3 protein which is regulated by post-translational mechanisms (ubiquitin degradation and tyrosine phosphorylation). The authors propose a "co-regulatory" model whereby the recruitment of BCAR3 to adhesions acts to modulate p130Cas tyrosine phosphorylation and cell migration. This manuscript would be of particular interest to cell and cancer biologists interested in the molecular regulation of cell migration.

(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 provides identification of different subpopulations of blood cells and gives new insights in putative hemocyte lineage relationships by single cell RNA sequencing. The main conclusions are fairly well supported by the data and this manuscript will be of high interest to crustacean immunologists and readers in the field of aquaculture.

(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 will be of interest to those in the fields of chromosome biology, mitotic regulation, and proteostasis. The authors put forward an interesting model of phosphodegron regulation of kinetochore assembly based on convincing genetic and biochemical data. The novel model will require some additional evidence before it can be considered well-supported, but the paper represents an advance in our knowledge of kinetochore regulation with experiments that are rigorous, well-designed and carefully conducted.

(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:

Oon and Prehoda report pulsatile contraction of apical membrane in the process of Par protein polarization in Drosophila neuroblasts. This explains how/why actin filament was required to localize/polarize Par complex. This very much resembles the observation in C. elegans embryos, and nicely unifies observations across systems.

(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:

Although the role of membrane potential in Cell Permeable Peptides (CPP) translocation has been consistently described in artificial systems, this multi scale study, combining cell biology, genetics and in silico approaches, further extends this topic to a live cell context where it shows that internalization stops when the membrane polarization is decreased by the removal of potassium channels. It proposes an original mechanism of CPP translocation based on water pore formation, which should be of interest for biophysicists, cell biologists and for applications such as drug delivery.

(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.)

Summary: The investigators' study of human AT2 cells-derived from induced pluripotent stem cells (iAT2 cells) in the presence and absence of a known dyskeratosis congenita (DC) pathogenic variant provides an exceptional model for understanding pathogenesis of pulmonary fibrosis in dyskeratosis congenita (DC) and related telomere biology disorders (TBDs). They provide convincing data demonstrating altered WNT signaling in iAT2 cells with short, dysfunctional telomeres and improved growth of iAT2 cells by GSK3 inhibition but fall short of convincingly showing the latter is due restored telomere end protection. The work should be of interest to those in the fields of telomere biology and the TBDs, lung physiology, WNT signaling and stem cell biology.

Evaluation Summary:

PKC-theta is known to regulate T cell activation, and this manuscript reveals a novel function of PKC-theta in the regulation of the nuclear pore complexes. The work by He and colleagues reveals that PKC-theta is recruited to the nuclear pore complex wherein it serves to regulate the assembly of key components of the RanBP2 subcomplex of the NPC, which in turn enables the translocation of AP1, NFkB and NFAT into the nucleus. However, these results need to be substantiated by additional experiments or by limiting the breath of the conclusions.

(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 describes the curation of a training dataset that will be an important resource for developers of new segmentation and deep-learning algorithms for electron microscopy data. The small size of the dataset makes it easy to use, and its broad range of image modalities ensure that the model will be applicable in many situations, making it very useful for the community.

(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 manuscript by Tindle et al describes generation of adult lung organoids (ALO) from human lung biopsies and their use to study the changes in gene expression as a result of SARS-CoV-2 infection. In this particular case the authors report the presence of AT1, AT2 cells, Basal cells, Goblet cells, Ciliated cells and Club cells. The authors were able to cultivate the cells at the air-liquid interface and to establish cultures of predominately proximal and predominately distal airway cells. The main finding is that proximal cells are more prone to viral infection, while distal cells are governing the exuberant inflammatory response, with both cells required for the exuberant response to occur. Useful information provided by the paper is the analysis gene signatures of various cellular models, in comparison to the infected human lung.

(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.)