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Validation of the cellular targets of microRNAs remains an ongoing priority. miR-CLIP, a new method based on psoralen crosslinking, immunoprecipitation and biotin affinity pulldowns, was applied to determine the miR-106a targetome, which included the H19 lncRNA.
Genetic evidence suggested jamABC from the jamaicamide biosynthetic pathway were responsible for the synthesis of the terminal alkyne functional group. Biochemical studies now confirm this activity and demonstrate the insertion of alkynes into two unrelated natural products.
Disulfide trapping and FRET studies define an agonist-induced conformational change in mGlu2 from inactive symmetric dimers with an interface at transmembrane domains (TMs) 4 and 5 to an active state with TM6s serving as the dimer interface.
Biocatalysis can take advantage of an enzyme's inherent reactivity regardless of its physiological role, as shown for a terpene cyclase turned Brønsted acid catalyst after its active site pocket was mutated while the activated aspartic acid was retained.
A family of cyclic lipopeptide natural products named lysocins was isolated from a soil bacteria sample and was found to exhibit antimicrobial actions. Genetic and biochemical evidence showed that lysocin E targets bacterial menaquinone.
An NMR structure reveals that the C terminus of the ubiquitin-conjugating enzyme Ube2w is disordered, leading to specific pairings with disordered substrates; loss of this sequence causes decreased substrate binding and ubiquitin transfer activity.
Inhibitors of FKBP51 with antidepressive activity are selective over the related FKBP52 and bind FKBP51 by an induced-fit mechanism that causes a conformational change. The analogous conformational change in FKBP52 generates a strained conformation.
A redox relay was identified in mammalian cells where the H2O2-reactive protein peroxiredoxin-2 oxidizes the transcription factor STAT3, resulting in the formation of transcriptionally inactive disulfide-linked oligomers.
Peptide natural product backbones are typically made ribosomally or by NRPS machinery. Exploration of pheganomycin biosynthesis defines a third hybrid model in which a grasp ligase joins an NRPS product with a ribosomally produced peptide.
The structure of complement regulatory protein factor H in complex with a preferred sialylated trisaccharide and the C3b thioester domain supports the idea of a ternary complex that mediates discrimination between self and nonself in a branch of innate immunity.
Bacterial MreB forms cytoskeletal filaments, rotating around the cell width, helping to shape cells. Imaging experiments indicate that MreB association with the B. subtilis cell membrane requires the peptidoglycan and wall teichoic acid precursor lipid II.
Biochemical, bioinformatic and genetic evidence uncover a tyrosine biosynthesis pathway in plants that—in contrast to known plant pathways—occurs in the cytosol, is insensitive to tyrosine feedback regulation and uses the traditionally bacterial prephenate dehydrogenase.
The loss of GSK-3 activity alters cellular responsiveness to kinase inhibitors such as mTOR and PLK1. A kinome-wide RNAi screen reveals that GSK-3 interacts with a third of known kinases.
A crystal structure of a chimera composed of lipid-metabolizing transmembrane enzymes HRASLS3 and LRAT (which catalyzes esterification of vitamin A) identifies a quaternary structural rearrangement that coincides with formation of a three-dimensionally swapped dimer.
Proteolysis must be carefully controlled to degrade desired targets without causing cellular damage. New data show that Lon protease in Enterobacteriaceae is regulated by a redox-dependent disulfide bond that determines the size of its exit pore.
The α1-adrenergic receptor antagonist terazosin protects flies and mammalian cells from stress and apoptosis through direct activation of the glycolytic enzyme phosphoglycerate kinase 1, which interacts with Hsp90 to promote ATP consumption.
β2-adrenergic receptor–mediated cAMP signaling from internal compartments is more effective compared to the plasma membrane in regulating a distinct set of genes, demonstrating a functional significance for the spatial separation of cAMP signaling.
Biosynthesis of the strigolactones—important plant hormones—has been solved up to carlactone. Biochemical and genetic evidence now demonstrate that homologous enzymes perform two subsequent oxidations, setting the strigolactone scaffold in place.
A collection of chemical tools and spectroscopic techniques demonstrate that Zn availability influences Cu+ storage and localization in the green alga Chlamydomonas, with Zn limitation causing the accumulation of Cu+ in lysosome-related organelles.
A modification of the in silico screening tool, DOCK, allows for identification of compounds that covalently modify catalytic and noncatalytic protein nucleophiles to modulate the activities of bacterial β-lactamase and the kinases RSK2, MSK1 and JAK3.