Biochemistry

Enhancing the protein fitness of interferon-lambda through computational design and glyco-engineering for prophylactic nasal drugs against respiratory viruses

1. Jeongwon Yun
2. Seungju Yang
3. Jae Hyuk Kwon
4. Luiz Felipe Vecchietti
5. Meeyoung Cha
6. Hyun Jung Chung
7. Ji Eun Oh
8. Ho Min Kim

Reviewed by Rapid Reviews Infectious Diseases

Concatenated modular BK channel constructs reveal divergent stoichiometry in gating control by LRRC26 (γ1), pore, and selectivity filter

1. Guanxing Chen
2. Qin Li
3. Kunal Shah
4. Jiusheng Yan

• Curated by eLife

  eLife Assessment

  In this important contribution, Yan and colleagues describe a powerful and compelling strategy to generate concatamers of the BK channel and their fusion constructs with the auxiliary gamma subunits, which allows exploring contributions of individual subunits of the tetrameric channel to its gating and the study of heteromeric channel complexes of defined composition. Distinct examples are presented, which illustrate great diversity in the stoichiometric control of BK channel gating, depending on the site and nature of molecular perturbations. The molecular approaches could be extended to other membrane proteins whose N and C termini face opposite sides of the membrane.

Reviewed by eLife

UV irradiation alters TFAM binding specificity and compaction of DNA

1. Dillon E King
2. Emily E Beard
3. Matthew J Satusky
4. Alex George
5. Ian Ryde
6. Caitlin Johnson
7. Emma L Dolan
8. Yuning Zhang
9. Wei Zhu
10. Hunter Wilkins
11. Evan Corden
12. Susan K Murphy
13. Dorothy A Erie
14. Raluca Gordân
15. Joel Meyer

• Curated by eLife

  eLife Assessment

  Mitochondrial DNA (mtDNA) exhibits a degree of resistance to mutagenesis under genotoxic stress, and this study on the mitochondrial Transcription Factor A (TFAM) presents important data concerning the possible mechanisms involved. The presented data are solid, technically rigorous, and consistent with established literature findings. The experiments are well-executed, providing convincing evidence on the change of TFAM-DNA interactions following UVC irradiation.

Reviewed by eLife

Cul5Wsb2 uses BCL2 proteins as co-receptors to target Bim for degradation

1. Wilhelm Vaysse-Zinkhöfer
2. Enya Marie Catherine Alcindor
3. Nicholas Garaffo
4. David Paul Toczyski

• Curated by eLife

  eLife Assessment

  The study presents a valuable finding on the ubiquitin-dependent regulatory loop in which proapoptotic Bim is targeted to the E3 ubiquitin ligase Cul5-Wsb2-mediated degradation through its sequestration by BCL2 proteins. The conclusions are supported by incomplete evidence and would benefit from additional experiments addressing both the mechanistic understanding and the physiological/cancer-related significance of the study.

Reviewed by eLife

Rare variant in intracellular loop-2 of the ghrelin receptor reveals novel mechanisms of GPCR biased signaling and trafficking

1. Elsa M. Balfe
2. Alexandre Torbey
3. Lara Kohlenbach
4. Jade A. Sency
5. Asuka Inoue
6. Lawrence S. Barak
7. Joshua D. Gross

Reviewed by PREreview

Multiple modes of cholesterol translocation in the human Smoothened receptor

1. Prateek D Bansal
2. Maia Kinnebrew
3. Rajat Rohatgi
4. Diwakar Shukla

• Curated by eLife

  eLife Assessment

  In this important study, the authors conducted extensive sets of computational and investigations of the mechanism of cholesterol transport in the smoothened (SMO) protein. The computational component integrated multiple state-of-the-art approaches such as adaptive sampling, free energy simulations, and Markov state modeling, providing compelling support for the proposed mechanistic model, which is further validated with solid experimental mutagenesis data.

Reviewed by eLife

Systematic characterization of site-specific proline hydroxylation using hydrophilic interaction chromatography and mass spectrometry

1. Hao Jiang
2. Jimena Druker
3. James W Wilson
4. Dalila Bensaddek
5. Jason R Swedlow
6. Sonia Rocha
7. Angus I Lamond

• Curated by eLife

  eLife Assessment

  The study presents a valuable resource of proline hydroxylation proteins for molecular biology studies in oxygen-sensing and cell signaling with the characterization of Repo-man proline hydroxylation site. The evidence supporting the claim of the authors is solid, although further clarification of the overall efficiency of the HILIC analysis, the specificity/sensitivity of immonium ion analysis, as well as quantification of proline hydroxylation identifications will be helpful. The work will be of interest to researchers studying post-translational modification, oxygen sensing, and cell signaling.

Reviewed by eLife

Determining the off-target activity of antibiotics and novel translation initiation sites in mitochondria

1. James Marks
2. Emma Young
3. Markus Hafner

• Curated by eLife

  eLife Assessment

  This valuable study presents findings on how prokaryotic antibiotics affect translation in mitochondrial ribosomes. Using mitoribosome profiling, the authors provide solid evidence that most tested antibiotics act similarly on bacterial and mitochondrial translation. Additionally, this work shows that alternative translation initiation events might exist in two specific mt-mRNAs (MT-ND1 and MT-ND5). However, additional biochemical and structural experiments are needed to support these findings.

Reviewed by eLife

Single-step in vitro ribosome reconstitution mediated by two GTPase factors, EngA and ObgE

1. Aya Sato
2. Weng Yu Lai
3. Yusuke Sakai
4. Yoshihiro Shimizu

• Curated by eLife

  eLife Assessment

  This is a useful study presenting solid data indicating that the bacterial GTPases EngA and ObgE enable single-step reconstitution of functional 50S ribosomal subunits under near-physiological conditions. The study elegantly bridges the gap between the non-physiological aspects of the previous two-step reconstitution method and the extract-dependent iSAT system to enable ribosome assembly under translation-compatible conditions; however, it is limited by reliance on rRNA and proteins extracted from native ribosomes and does not achieve a true bottom-up reconstruction from all synthetic components. The evidence is incomplete in not characterizing the spectrum of reporter polypeptides produced and not comparing their rate and yield of synthesis from reconstituted ribosomes to that obtained with pure native ribosomes; and the impact of the study is limited by not including reporters to examine the fidelity of initiation, elongation or termination achieved with the reconstituted ribosomes.

Reviewed by eLife

Automated genome mining predicts structural diversity and taxonomic distribution of peptide metallophores across bacteria

1. Zachary L Reitz
2. Bita Pourmohsenin
3. Melanie Susman
4. Emil Thomsen
5. Daniel Roth
6. Alison Butler
7. Nadine Ziemert
8. Marnix H Medema

• Curated by eLife

  eLife Assessment

  This important and compelling study establishes a robust computational and experimental framework for the large-scale identification of metallophore biosynthetic clusters. The work advances beyond current standards, providing theoretical and practical value across microbiology, bioinformatics, and evolutionary biology.

Reviewed by eLife