Molecular Biology

A cryogenic, coincident fluorescence, electron, and ion beam microscope

1. Daan B Boltje
2. Jacob P Hoogenboom
3. Arjen J Jakobi
4. Grant J Jensen
5. Caspar TH Jonker
6. Max J Kaag
7. Abraham J Koster
8. Mart GF Last
9. Cecilia de Agrela Pinto
10. Jürgen M Plitzko
11. Stefan Raunser
12. Sebastian Tacke
13. Zhexin Wang
14. Ernest B van der Wee
15. Roger Wepf
16. Sander den Hoedt

• Curated by eLife

  eLife assessment

  This paper is of particular interest to researchers who plan to use focused-ion beam scanning electron microscopes (FIB-SEMs) and require fluorescent data to guide the milling process. The authors describe a valuable after-market upgrade that allows fluorescent data acquisition during FIB-milling without stage repositioning. Technical details of the fluorescent module upgrade together with the sample stage redesign are compellingly documented and will enhance the implementation of this important technology.

Reviewed by eLife

The anatomy of transcriptionally active chromatin loops in Drosophila primary spermatocytes using super-resolution microscopy

1. Madeleine L. Ball
2. Stefan A. Koestler
3. Leila Muresan
4. Sohaib Abdul Rehman
5. Kevin O’Holleran
6. Robert White

Reviewed by Review Commons

Structure and flexibility of the yeast NuA4 histone acetyltransferase complex

1. Stefan A Zukin
2. Matthew R Marunde
3. Irina K Popova
4. Katarzyna M Soczek
5. Eva Nogales
6. Avinash B Patel

• Curated by eLife

  eLife assessment

  This manuscript provides insights into the architecture of the yeast histone acetyltransferase complex NuA4 and is of broad interest to those studying transcription and chromatin modification. The cryo-EM data are of very high quality, and enable the authors to devise a structural model that is in much better agreement with biochemical data than previously reported models. This structure represents an important puzzle piece towards a molecular understanding of chromatin modification.

Reviewed by eLife

Nuclear m6A reader YTHDC1 promotes muscle stem cell activation/proliferation by regulating mRNA splicing and nuclear export

1. Yulong Qiao
2. Qiang Sun
3. Xiaona Chen
4. Liangqiang He
5. Di Wang
6. Ruibao Su
7. Yuanchao Xue
8. Hao Sun
9. Huating Wang

• Curated by eLife

  eLife assessment

  This is a valuable study performing elegant experiments making identification of a specific regulator in skeletal muscle regeneration. It will form a foundation for further mechanistic investigation. The work is of importance in the clinical field of muscle injury and regeneration.

Reviewed by eLife

Tup1 is critical for transcriptional repression in Quiescence in S. cerevisiae

1. Thomas B. Bailey
2. Phaedra A. Whitty
3. Eric U. Selker
4. Jeffrey. N. McKnight
5. Laura E. McKnight

Reviewed by Review Commons

A cell wall synthase accelerates plasma membrane partitioning in mycobacteria

1. Takehiro Kado
2. Zarina Akbary
3. Daisuke Motooka
4. Ian L Sparks
5. Emily S Melzer
6. Shota Nakamura
7. Enrique R Rojas
8. Yasu S Morita
9. M Sloan Siegrist

• Curated by eLife

  eLife assessment

  This paper addresses an important question: the relationship between the cell wall and other, primarily lipid, based components of the cell envelope. Building on previous work, the authors provide data suggesting that the activity of a PonA2, non-essential peptidoglycan synthase, promotes membrane partitioning through its role in cell wall synthesis. While the data are consistent with this model, the reviewers felt additional experiments are necessary to fully support the authors' conclusions.

Reviewed by eLife

Perturbed fatty-acid metabolism is linked to localized chromatin hyperacetylation, increased stress-response gene expression and resistance to oxidative stress

1. Jarmila Princová
2. Clàudia Salat-Canela
3. Petr Daněk
4. Anna Marešová
5. Laura de Cubas
6. Jürg Bähler
7. José Ayté
8. Elena Hidalgo
9. Martin Převorovský

Reviewed by Review Commons

Molecular mechanism of Afadin substrate recruitment to the receptor phosphatase PTPRK via its pseudophosphatase domain

1. Iain M Hay
2. Katie E Mulholland
3. Tiffany Lai
4. Stephen C Graham
5. Hayley J Sharpe
6. Janet E Deane

• Curated by eLife

  Evaluation Summary:

  These studies establish a role for the D2 pseudophosphatase domain of the PTPRK receptor-like phosphotyrosine phosphatase in recruiting Afadin, a cell-cell junction protein that is reported to be a PTPRK substrate, for dephosphorylation by the active D1 phosphatase domain. These findings suggest that the D2 pseudophosphatase domains of RPTPKs might have a general function as platforms to recruit specific phosphotyrosine substrates.

  (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 #3 agreed to share their name with the authors.)

Reviewed by eLife

Development of a versatile high-throughput mutagenesis assay with multiplexed short-read NGS using DNA-barcoded supF shuttle vector library amplified in E. coli

1. Hidehiko Kawai
2. Ren Iwata
3. Shungo Ebi
4. Ryusei Sugihara
5. Shogo Masuda
6. Chiho Fujiwara
7. Shingo Kimura
8. Hiroyuki Kamiya

Reviewed by Review Commons

Inhibited KdpFABC transitions into an E1 off-cycle state

1. Jakob M Silberberg
2. Charlott Stock
3. Lisa Hielkema
4. Robin A Corey
5. Jan Rheinberger
6. Dorith Wunnicke
7. Victor RA Dubach
8. Phillip J Stansfeld
9. Inga Hänelt
10. Cristina Paulino

• Curated by eLife

  Evaluation Summary:

  KdpFABC is a bacterial potassium uptake transporter made up of a channel-like subunit (KdpA) and a P-type ATPase (KdpB). When potassium levels are low (< 2 mM), the transporter actively and selectively uptakes potassium, but must be switched off again to prevent excessive K+ accumulation. Although structures of KdpFABC have been determined before, the structural basis for inhibition by phosphorylation is unknown. Here, the authors have determined the structure of KdpABC in an arrested (off-state) that is in a distinct conformation from previously determined P-type ATPase structures. More detailed structural comparisons are needed to more convincingly show this, however, and the protein required to inhibit KdpABC by phosphorylation remains unknown. This paper will be of interest to researchers in the microbiology and transporter communities.

  (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