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The incorporation of nitrogen in steroidal glycoalkaloids is hypothesized to occur through a transamination reaction. Here, the authors show that GLYCOALKALOID METABOLISM12 appears to evolve from the canonical γ-aminobutyric acid transaminases and directs the biosynthesis of nitrogen-containing steroidal metabolites in Solanum plants.
Caliendo and Vitu et al. developed a platform for endogenous gene expression monitoring and conditional actuator activation, by integrating noncoding RNAs downstream of the polyadenylation signal without altering endogenous gene coding sequences.
Superoxide oxidizes the iron–sulfur cluster in the DNA demethylase ROS1 and extensively participates in the establishment of active DNA demethylation in plants to maintain stem cell fate.
By enriching productive mutational paths, a Kemp eliminase that speeds up proton abstraction >108-fold was developed in only five evolution rounds. Recombining it with a variant differing by 29 substitutions revealed the underlying fitness landscape.
How a lasso cyclase ties a lasso peptide into its characteristic knot has remained poorly understood. Here the authors identify key molecular interactions that guide lasso peptide folding and cyclase substrate tolerance to inform cyclase engineering for expanded lasso peptide diversity.
Cryo-electron microscopy and biochemical analysis reveal the activation mechanism of protein arginine kinase McsB by its activator McsA for protein quality control under stress in Gram-positive bacteria.
Development of a ligand-based proximity labeling strategy enables interrogation of the native cell membrane interactome for the GLP-1 receptor in two cell types, revealing new regulators of receptor-mediated signaling and β cell responses.
A method to study G-protein-coupled receptor (GPCR) trafficking has been developed using engineered APEX2 and CRISPR interference screening. The innovative approach reveals a network of proteins coordinated by DNAJC13 that control efficient GPCR sorting into degradative or recycling pathways.
Development of a biosensor for GPCR trafficking to the lysosome combined with a genome-wide CRISPR screen identified DNAJC13 as a critical regulator of agonist-induced trafficking of the δ-opioid receptor to the lysosome.
Qiao and Nguyen et al. describe a strategy to block hypoxia-dependent gene expression in cell and mouse models of breast cancer through dual targeting of X-box-binding protein 1 and hypoxia-inducible factor binding to DNA with fully synthetic, stabilized artificial transcription factors.
Cao et al. introduce artificial lncRNA (alncRNA) for targeted protein degradation by engineering and linking lncRNA HOTAIR sequence to RNA aptamers. The resulting alncRNAs degrade oncogenic proteins, such as c-MYC and KRAS, showing promise for cancer therapies.
Pan et al. establish a general photoproximity labeling approach in living cells, applying microenvironment mapping with a HaloTag-based platform (HaloMap) to profile the stress granule proteome and identify ubiquitin-related modulators as key mediators of granule disassembly.
Organisms regulate biogenic crystal properties for various functions. Here the authors reveal the genetic and biochemical control of crystal morphogenesis in zebrafish iridophores, showing that the chemical composition, influenced by enzyme expression, determines crystal morphology and functionality.
Engineered demethylase LSD1 opens a new avenue in developing tools to study intricate relationships between histone post-translational modifications that can be enzymatically edited.
A method called massively parallel PPI measurement by sequencing (MP3-seq) is developed for measuring protein–protein interactions at scale. MP3-seq uses DNA barcodes that are associated with specific protein pairs and provides a quantitative measure of interaction strength. Interactions between rationally designed heterodimers and elements conferring interaction specificity were investigated using MP3-seq.
Assembly-line polyketide synthases perform modular enzymatic synthesis of medically important natural products. In this study, the authors used site-selective crosslinking to probe structural states that reveal how substrate carrier proteins interact with successive modules in a representative assembly line.
Spores harboring the gene circuit for the secretory expression of Burkholderia cepacia lipase were processed with poly(caprolactone) pellets to manufacture living plastics. Spore incorporation did not compromise the properties of the materials. Damage to the plastic surface and triggering of germinated cells caused secretion of the lipase, leading to depolymerization.
