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Förster resonance energy transfer (FRET) is essential for studying molecular interactions, yet challenges remain in mapping complex interactions in cellular environments. Here, the authors employ a three-fluorophore FRET-cascade system with time-correlated single photon counting fluorescence lifetime imaging microscopy to elucidate Rap1-interacting adaptor molecule (RIAM)-vinculin interactions in focal adhesions, revealing mechanosensitive roles of vinculin and its interaction with RIAM in a force-independent manner.

The combination of single-molecule fluorescence detection with Förster resonance energy transfer provides a powerful probe of biomolecular dynamics on timescales ranging from nanoseconds to days. This Review outlines single-molecule Förster resonance energy transfer spectroscopy with a focus on dynamics and highlights future developments and enhanced capabilities.

Lead sulphide colloidal nanocrystals are now used to harvest non-emissive triplet excitons generated in a tetracene layer. Depending on the length of the ligands capping the nanocrystals, exciton transfer efficiency as high as 90% can be achieved.

Fair climate targets aligned with the Paris Agreement can be calculated in multiple ways, yielding diverse outcomes. Researchers unpack how equity, global strategies and political and social uncertainties shape fair share allocations, using them to assess nationally determined contributions and guide global climate finance.

How chemotherapeutic nucleoside 6-thio-2’-deoxyguanosine (6-thiodG) targets telomerase to inhibit telomere maintenance in cancer cells and tumors was unclear. Here, the authors show that telomere length and telomerase status determine 6-thio-dG sensitivity and uncover the molecular mechanism by which 6-thio-dG selectively inhibits telomerase synthesis of telomeric DNA.

Development of the bright green and red fluorescent proteins, Clover and mRuby2, creates a fluorescence resonance energy transfer (FRET) pair with the highest Förster radius among existing ratiometric FRET pairs. Substitution of this pair for current FRET pairs in several existing sensors reliably and substantially improves sensor performance.

By combining single-molecule spectroscopy, nanophotonic enhancement, and molecular simulations, the authors reveal the extremely rapid chain dynamics of single-stranded nucleic acids.

Variants in the PSMC5 gene impair proteasome function and cellular homeostasis, altering brain development in children. This study reveals underlying molecular mechanisms contributing to this neurodevelopmental phenotype, and suggests therapeutic leads for neurodevelopmental proteasomopathies.

A multi-laboratory study finds that single-molecule FRET is a reproducible and reliable approach for determining accurate distances in dye-labeled DNA duplexes.

A fresh approach to protein design that incorporates excited intermediate states enables precise control over the lifetime of protein interactions, with potential applications in cell-signalling modulation and in biosensors and synthetic circuits.

Programming the 3D spatial organization of quantum dots requires precise control over their individual valence, but this is challenging due to the possible presence of multiple binding sites. Here, authors develop a general approach that uses highly programmable wireframe DNA origami structures to control the 3D spatial relationships between QDs and other non-nucleic-acid molecules.

Large-scale commercialization of organic light-emitting diodes is impeded by the short operational lifetime of blue emitting materials. Leeet al. show a strategy to manage the energy dissipation on molecular dissociation using dopants with high triplet exciton energy that improves device stability.

High-depth sequencing of non-cancerous tissue from patients with metastatic cancer reveals single-base mutational signatures of alcohol, smoking and cancer treatments, and reveals how exogenous factors, including cancer therapies, affect somatic cell evolution.

Hepatitis C virus (HCV) variability and its phenotypic consequences aren’t well studied in relation to viral replication fitness and disease severity. Here, the authors identify a replication-enhancing domain in non-structural protein 5A, linking high replication fitness to severe disease outcomes, with implications for understanding HCV pathogenesis in immunocompromised patients.

Two highly charged disordered human proteins phase-separate into viscous complex coacervates while retaining their rapid conformational dynamics through pico- to nanosecond exchange of short-lived side-chain interactions.

T4 Lysozyme (T4L) is a model protein whose structure is extensively studied. Here the authors combine single-molecule and ensemble FRET measurements, FRET-positioning and screening and EPR spectroscopy to study the structural dynamics of T4L and describe its conformational landscape during the catalytic cycle by an extended Michaelis–Menten mechanism and identify an excited conformational state of the enzyme.

The development of antimalarials against the human liver and asexual blood stages is one of the top public health challenges. Here, the authors report a single-step biochemical assay for the characterization of prolyl-tRNA synthetase inhibitors, and develop high-affinity inhibitors for the enzyme, including elusive triple-site ligands.

Evidence suggests oligomerisation of G protein-coupled receptors in membranes, but this is controversial. Here, authors use single-molecule and ensemble FRET, and spectroscopy to show that the neurotensin receptor 1 forms multiple dimer conformations that interconvert - “rolling” interfaces.

Nanocrystal quantum dots are excellent emitters of light, but energizing that emission electrically has proved difficult. A newly discovered non-radiative energy-transfer process that allows electron–hole pairs to be generated in quantum wells, and then transferred into quantum dots, could lead to a new generation of ultra-high-efficiency light sources for use at scales ranging from nano-optics to a city block.

Cyan variants of green fluorescent protein (CFPs) are widely used as donors in FRET experiments. Here, a new CFP, mTurquoise2, is developed, which displays a high-fluorescence quantum yield and a long mono-exponential fluorescence lifetime.

HIV-1 replication is inhibited by the enzyme APOBEC3G via two separate mechanisms. A deamination mechanism requires rapid binding and release of single-stranded DNA (ssDNA), whereas a roadblock mechanism requires slow binding. Now APOBEC3G has been shown to initially bind ssDNA with rapid on–off rates. The enzyme subsequently converts via oligomerization to a slowly dissociating binding mode, which, it is proposed, inhibits reverse transcription.

Bacterial divisome protein FtsA, which is an actin homologue, forms double filaments following binding to FtsN, and like MreB, an actin homologue in the elongasome, the curvature-sensing double filaments guide peptidoglycan insertion for cell division.

Here, the authors compare HP1 from fission yeast, fly and mouse, and find that the propensity of HP1 to phase-separate and to cluster heterochromatin decrease in this order, suggesting an evolutionary adaptation of HP1 function.

Strongly interacting dye molecule complexes with coherent energy transport can be organized through programmable DNA scaffolds.

Through careful management of the bound electron–hole pairs responsible for light emission, efficiencies can be achieved that should challenge those of the incandescent sources responsible for a substantial amount of domestic and commercial electricity consumption.

The association and dissociation dynamics of the ECF transporter complex for vitamin B12 are visualized by single-molecule FRET, highlighting the original transport mechanism of this group of ABC transporters.

CoraFluors, a class of macrocyclic terbium complexes for use in time-resolved FRET, exhibit physicochemical properties desirable for biological studies, including characterization of Keap1 ligands and HDAC1 target engagement profiling in live cells.

A consensus cell type atlas for the fly brain provides both an intellectual framework and open-source toolchains for brain-scale comparative connectomics.

SIRT6 plays essential roles in metabolism, tumor suppression, and DNA repair through the deacetylation of histone substrates. Here the authors use biophysical methods to investigate the molecular basis for SIRT6 interaction with the nucleosome core particle.

Fatty liver disease exacerbates atherosclerosis, but the underlying mechanisms remain unclear. Here, the authors show that D-dopachrome tautomerase (D-DT/MIF-2) acts as an atypical chemokine, promoting both atherosclerosis and hepatic lipid accumulation.

Here, Forster et al. compare 1354 cultured commensal strains (540 species) to 45,403 pathogen strains (12 species), identifying 64,188 MGE-mediated antibiotic resistance gene transfer events between the two groups, and show that 15 broad host range MGEs are able to transfer between phyla.

Photon upconversion methods demonstrated thus far involve challenging requirements. Here Weingartenet al. demonstrate a mechanism called cooperative energy pooling, in which multiple photoexcited sensitizers resonantly and simultaneously transfer their energies to a higher-energy state on a single acceptor.

Ataxia telangiectasia and Rad3-related kinase (ATR) is a rational radiosensitization target. This trial evaluates the combination of the ATR inhibitor, ceralasertib, with palliative radiotherapy in 27 patients with advanced solid tumors.

Bacteria represent an unexploited reservoir of biosensing proteins. Here the authors use genomic screens and functional assays to isolate a progesterone sensing allosteric transcription factor and use a FRET-based method to develop an optical progesterone sensor.

The presence of conformational substates of a catalytically competent 'closed' state in the ligand-free form of adenylate kinase is detected. Molecular dynamics simulations indicated that the partially closed conformations were sampled in nanoseconds, and NMR and single-molecule FRET experiments revealed the sampling of a fully closed conformation occurring on the microsecond-to-millisecond timescale.

Human proteins SERINC3 and SERINC5 are HIV-1 restriction factors that reduce viral infectivity. Here, the authors show that SERINC3 has architecture resembling non-ATP dependent lipid transporters and induces loss of membrane asymmetry correlated with changes in envelope conformation and loss of infectivity.

Morphogenesis of tissue sheets is well studied, but mechanisms that shape bulk tissues are unclear. Here, the authors show that mesenchymal cells intercalate in 3D to shape the mouse branchial arch, with cortical forces driving intercalations in a Wnt5a-, Yap/Taz- and Piezo1-dependent manner.

Rudalska et al. describe a novel class of p38α inhibitors with increased target residence time. They explore the drugs’ specificity, pharmacokinetics and toxicity profile and show that they are efficacious in the context of colorectal cancer.

Integrin β₂ attachment regulates inflammation via effects on neutrophil rolling and extravasation through sequential integrin extension then headpiece opening. Here the authors show an alternative open headpiece prior to extension stabilized in cisby ICAM-1 that limits neutrophil adhesion.

STIM proteins are key regulators of intracellular Ca²⁺ signalling. Here, Wang et al. demonstrate that subtle differences between STIM1 and its close homologue STIM2 have profound consequences for their ability to gate Orai1 Ca²⁺channels, thus revealing the basis for their distinct physiological functions.

Combination of epidemiology, preclinical models and ultradeep DNA profiling of clinical cohorts unpicks the inflammatory mechanism by which air pollution promotes lung cancer

Coherence observed in chemical and biological systems suggests that even in the presence of disorder and noise the phenomenon may yield transformative ways for improving function.

NanoLuc substrates with improved solubility and bioavailability, hydrofurimazine and fluorofurimazine, strongly enhance bioluminescence signals in vivo and enable bright dual-color bioluminescent imaging with AkaLuc and AkaLumine.

Temperature jump technique is widely used to probe the fast dynamics of protein and DNA folding, but constrained to modest temperature control. Here, the authors use a microfluidic device combined with an infrared laser to heat or cool DNA hairpins up to 70 °C on a microsecond time scale.

High data volumes from multidimensional imaging techniques can lead to slow collection and processing times. Here, the authors implement multispectral fluorescence lifetime imaging microscopy (FLIM) that uses time-correlated photon counting technology to reach simultaneously high imaging rates combined with high spectral and temporal resolution.