Cell Biology

Disrupting the ciliary gradient of active Arl3 affects rod photoreceptor nuclear migration

1. Amanda M Travis
2. Samiya Manocha
3. Jason R Willer
4. Timothy S Wessler
5. Nikolai P Skiba
6. Jillian N Pearring

• Curated by eLife

  Evaluation Summary:

  The current paper is of interest to cell biologists studying ciliogenesis and specifically vertebrate photoreceptors, which are specialized cilia. The study shows that mutations in the small GTP binding protein ARL3 known to cause dominant inherited retinal dystrophies in humans result in ARL3 hyperactivity, disrupting the normal ciliary gradient of ARL3 activity and leading to altered retinal development. The authors demonstrate restored normal nuclear distribution by overexpression of ARL3 effectors, suggesting that the active mutants disrupt nuclear migration, at least in part, by sequestering ARL3 effectors. Overall, the experiments are properly controlled, executed, and analyzed and involve a series of extensive biochemical analyses complemented with in vivo phenotypic assessment. The development of a method to analyze snapshots of the interaction between ARL3 and its interactors is also a strength of the paper, however, significant concerns remain regarding links between nuclear migration failure and ciliogenesis in the outer segment, and alternative possibilities that could explain the phenotype of the ARL3 Y90C mutant with respect to its sequestration of the GEF ARL13B. Addressing these major concerns would improve the manuscript and could have considerable impact on the field.

  (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.)

Reviewed by eLife

Short-term molecular consequences of chromosome mis-segregation for genome stability

1. Lorenza Garribba
2. Giuseppina De Feudis
3. Valentino Martis
4. Martina Galli
5. Marie Dumont
6. Yonatan Eliezer
7. René Wardenaar
8. Marica Rosaria Ippolito
9. Divya Ramalingam Iyer
10. Andréa E. Tijhuis
11. Diana C. J. Spierings
12. Michael Schubert
13. Silvia Taglietti
14. Chiara Soriani
15. Simon Gemble
16. Renata Basto
17. Nick Rhind
18. Floris Foijer
19. Uri Ben-David
20. Daniele Fachinetti
21. Ylli Doksani
22. Stefano Santaguida

Reviewed by ASAPbio crowd review

lncRNA H19/Let7b/EZH2 axis regulates somatic cell senescence

1. Manali Potnis
2. Justin Do
3. Olivia El Naggar
4. Eishi Noguchi
5. Christian Sell

• Curated by eLife

  eLife Assessment

  In this manuscript, Sell et al., investigate the role of the long non-coding RNA H19 in regulating cellular senescence. Using several cell models they identify upstream and downstream effectors of H19 including let-7 and EZH2. The advances in this work include the identification of a specific cascade of factors connecting H19, senescence and the actions of rapamycin.

Reviewed by eLife, ASAPbio crowd review

Wolbachia action in the sperm produces developmentally deferred chromosome segregation defects during the Drosophila mid-blastula transition

1. Brandt Warecki
2. Simon William Abraham Titen
3. Mohammad Shahriyar Alam
4. Giovanni Vega
5. Nassim Lemseffer
6. Karen Hug
7. Jonathan S Minden
8. William Sullivan

• Curated by eLife

  Evaluation Summary:

  This manuscript describes cellular and developmental defects at late embryonic stages during Wolbachia-induced cytoplasmic incompatibility (CI), which occurs when male insects harboring the endosymbiont bacteria Wolbachia fertilize eggs of uninfected females, triggering embryonic lethality, usually at the first nuclear division. This work presents evidence that the mechanism of late embryonic defects is independent from the ones responsible for early embryonic defects. The experiments are technically superb, and the strength of evidence provided is compelling, including beautiful single-embryo PCR analyses and convincing light microscopy. While the overall significance might be limited, the knowledge will be useful to those in the fields of cytoplasmic incompatibilities and insect embryo development.

  (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.)”

Reviewed by eLife

Notch signaling functions in noncanonical juxtacrine manner in platelets to amplify thrombogenicity

1. Susheel N Chaurasia
2. Mohammad Ekhlak
3. Geeta Kushwaha
4. Vipin Singh
5. Ram L Mallick
6. Debabrata Dash

• Curated by eLife

  Evaluation Summary:

  Advances in the discovery of novel anti-platelet therapeutics remains an unmet need. This manuscript by Chaurasia et al. describes a novel signaling pathway involving Notch1 and its ligand, Delta-like ligand-4 (DLL4) in driving platelet activation and thrombus formation. The authors provide convincing mechanistic studies to show that blockade of this pathway may serve as a new therapeutic approach to prevent/treat thrombosis. The work will be of great interest to individuals in the hematology and thrombosis field.

  (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.)

Reviewed by eLife

PLK4 drives centriole amplification and apical surface area expansion in multiciliated cells

1. Gina M LoMastro
2. Chelsea G Drown
3. Aubrey L Maryniak
4. Cayla E Jewett
5. Margaret A Strong
6. Andrew Jon Holland

• Curated by eLife

  Evaluation Summary:

  PLK4 is the master regulator of centriole biogenesis, but whether it is also key for centriole amplification during differentiation of multiciliated cells (MCCs) has been questioned based on PLK4 chemical inhibition. Here, using mouse models engineered to lack PLK4 or PLK4 activity, LoMastro et al provide very compelling evidence that PLK4 and its activity are essential for centriole amplification in MCCs. Moreover, they show that centriole amplification in MCCs drives expansion of their apical surface. The findings will be of interest to cell biologists and experts interested in multi-ciliogenesis-related pathologies.

  (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.)

Reviewed by eLife

Crosshair, semi-automated targeting for electron microscopy with a motorised ultramicrotome

1. Kimberly Meechan
2. Wei Guan
3. Alfons Riedinger
4. Vera Stankova
5. Azumi Yoshimura
6. Rosa Pipitone
7. Arthur Milberger
8. Helmuth Schaar
9. Inés Romero-Brey
10. Rachel Templin
11. Christopher J Peddie
12. Nicole L Schieber
13. Martin L Jones
14. Lucy Collinson
15. Yannick Schwab

• Curated by eLife

  Evaluation Summary:

  Meechan et al. present a systematic approach to semi-automate an ultramicrotome operation for targeting a specific plane aided by x-ray tomography measurements. It is a fundamental work of great interest to any users of using electron microscopy (EM), particularly when targeting the imaging of thin sections in a select region of interest by ultramicrotomy, or when targeting volume EM of select sample regions. The manuscript documents with exceptional detail a workflow including both microtome modifications and software adaptations for semi-automated targeting of structures with micrometer precision, resulting in a faster and more accurate orientation of the image acquisition planes for volume electron microscopy, a task that has traditionally been difficult and time-consuming. Therefore, this work will reduce sample preparation labor and, critically, facilitate the comparison of the ultrastructure of multiple samples. The method is based on X-ray imaging acquisition prior to any sectioning and proposes a solution for the two instruments commercially available in the field, and by transparently sharing all the data, hardware, and software, and by describing every detail of the workflow, this fundamental method can be readily adopted by any practitioner, enabling its wide application - it is a key step in the field regarding speed-up, accuracy, and reproducibility in electron microscopy.

  (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 names with the authors.)

Reviewed by eLife

Nanog organizes transcription bodies

1. Ksenia Kuznetsova
2. Noémie M. Chabot
3. Martino Ugolini
4. Edlyn Wu
5. Manan Lalit
6. Haruka Oda
7. Yuko Sato
8. Hiroshi Kimura
9. Florian Jug
10. Nadine L. Vastenhouw

Reviewed by ASAPbio crowd review

Adaptation to glucose starvation is associated with molecular reorganization of the circadian clock in Neurospora crassa

1. Anita Szőke
2. Orsolya Sárkány
3. Géza Schermann
4. Orsolya Kapuy
5. Axel CR Diernfellner
6. Michael Brunner
7. Norbert Gyöngyösi
8. Krisztina Káldi

• Curated by eLife

  Evaluation Summary:

  This manuscript will be of interest to researchers working in chronobiology and metabolism. The authors have found evidence that starvation decreases the abundance of the fungal circadian clock protein white collar complex (WCC), even though WC-1 is required for responses to starvation. This observation is interesting, but the authors should consider that WCC has several other functions (as a light receptor, in transcriptional regulation) that are not necessarily clock connected. As such the most interesting result from this paper is that the standard model for the molecular mechanism of the fungal circadian clock does not explain the persistence of normal rhythms under extreme starvation conditions, where the levels of clock proteins are drastically altered.

  (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.)

Reviewed by eLife

β-cell deletion of the PKm1 and PKm2 isoforms of pyruvate kinase in mice reveals their essential role as nutrient sensors for the KATP channel

1. Hannah R Foster
2. Thuong Ho
3. Evgeniy Potapenko
4. Sophia M Sdao
5. Shih Ming Huang
6. Sophie L Lewandowski
7. Halena R VanDeusen
8. Shawn M Davidson
9. Rebecca L Cardone
10. Marc Prentki
11. Richard G Kibbey
12. Matthew J Merrins

• Curated by eLife

  **Evaluation Summary:
  **
  This manuscript employs in vitro studies and elegant mouse models to detail how specific pyruvate kinase isoforms impact pancreatic beta-cell ATP/ADP levels, ATP-sensitive K+ channel (KATP channel) activity, calcium handling, and insulin secretion. This is an important study that challenges the current paradigms of KATP-channel regulation, the major signaling mechanism through which pancreatic beta cells couple blood glucose levels to insulin release. Future studies will be necessary to determine whether similar mechanisms are used in human pancreatic beta cells.

  (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 and Reviewer #3 agreed to share their names with the authors.)

Reviewed by eLife