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A chemoproteomic strategy called RNA-mediated activity-based protein profiling (RNABPP) was developed to profile RNA-modifying enzymes in live cells and uncover the RNA substrates of the human dihydrouridine synthase DUS3L.
Structures and biochemistry reveal how covalent linkage of a ubiquitin-like protein elicits protein–protein interactions indirectly. NEDD8 allosterically activates CUL5-RBX2 cullin-RING E3 binding to the ARIH2 RBR-type E3 for joint ubiquitylation.
The development of a photocage-nanobody based technology enabled in-depth analysis of live cells from tissues while retaining their spatial information.
Structural and biochemical analysis of the plesiocin macrocyclase enzyme PsnB revealed that, unlike other ribosomal natural products, the core region of the precursor peptide enhances its interaction with the enzyme via conserved glutamate residues.
Bifunctional ‘MoDE-A’ molecules, which contain ligands that bind to an extracellular protein and carbohydrate residues that recruit it to the asialoglycoprotein receptor, mediate cellular uptake and lysosomal turnover of target proteins.
Structures of three cyanophycin synthetases reveal how the constituent glutathione synthetase and muramyl ligase-like domains cooperate to make cyanophycin, a poly-aspartate chain with arginine residues attached to the sidechains by isopeptide bonds.
Deploying two unrelated CRISPR nucleases in tandem, with multiplexed CRISPR RNAs and a chemically stabilized activator, creates a simple, one-step assay that can rapidly detect attomolar concentrations of RNA without needing target amplification.
Controlled assembly of synthetic intracellular condensates from engineered disordered proteins enables control of cellular activities such as proliferation and division through recruitment, sequestration and insulation of endogenous client proteins.
The cystic fibrosis transmembrane conductance regulator anion channel can adopt an alternate conformation of its nucleotide-binding domain, which affects channel activity and, under certain conditions, leads to unfolding and protein degradation.
Approximately half of lipoproteins destined to the Escherichia coli outer membrane display intrinsically disordered linker peptides at their N termini, which are required for optimal trafficking by the Lol lipoprotein sorting system.
Combined use of microcrystal electron diffraction and genome mining for biosynthetic gene clusters enables the rapid structural elucidation of natural products, including a newly discovered 2-pyridone compound and a revised structure of fischerin.
The full-length structure of HUWE1 reveals the bipartite organization of a giant E3 ubiquitin ligase, comprising a catalytic HECT domain and a large, ring-shaped scaffold that provides docking sites for various substrates and regulates E3 activity.
Collagen cross-linking, mediated by lysyl oxidases (LOX), is critical for the stability of the extracellular matrix. Aldehyde-reactive sensors and collagen peptide probes for monitoring LOX activity enable direct in vivo imaging of collagen maturation.
A knot-like RNA from Zika virus is shown to evade digestion by host RNases through its extreme resistance to mechanical unfolding. Weakening the knot mechanically or reducing the likelihood of knot formation lowers the RNase resistance.
A linkage-specific tool for K29-linked polyubiquitin was developed, enabling the discovery that K29-linked ubiquitination participates in multiple cellular pathways, including the proteotoxic stress response and cell cycle regulation.
Design of a bivalent inhibitor containing an ATP-competitive moiety and rapamycin-modified FRB binding ligand that selectively inhibits mTORC1 results in potent and durable inhibition of 4EBP1 phosphorylation and cell proliferation in vitro and in vivo.
Autonomous hypermutation yeast surface display (AHEAD) mimics the process of somatic hypermutation in animals to enable the rapid in vitro evolution of antibodies, including nanobodies targeting the RBD of SARS-CoV-2.
Degradation-tuning RNAs (dtRNAs) have the capacity to increase and decrease RNA stability in vivo and in vitro. Appending these modules to transcripts allows fine tuning of circuit dynamics, CRISPR interference and paper-based viral diagnostics.
Fragment screening and medicinal chemistry optimization led to development of a small-molecule inhibitor of RING1B–BMI1 E3 ligase, blocking the H2A ubiquitination activity of the Polycomb repressive complex 1 and inducing differentiation in leukemia cells.