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This study identified the LARP6 La Module from Tetrabaena socialis (T. socialis), a four-celled green algae, in an effort to better understand the evolution of LARP6 structure and RNA-binding activity in multicellular eukaryotes. Using a combination of sequence alignments, domain boundary screens, and structural modelling, we recombinantly expressed and isolated the TsLARP6 La Module to > 98% purity for in vitro biochemical characterization. The La Module is stably folded and exerts minimal RNA binding activity against single-stranded homopolymeric RNAs. Surprisingly, it exhibits low micromolar binding affinity for the vertebrate LARP6 cognate ligand, a bulged-stem loop found in the 5'UTR of collagen type I mRNA, but does not bind double-stranded RNAs of similar size. These result suggests that the TsLARP6 La Module may prefer structured RNA ligands. In contrast, however, the TsLARP6 La Module does not exhibit the RNA chaperone activity that is observed in vertebrate homologs. Therefore, we conclude that protist LARP6 may have both distinct RNA ligands and binding mechanisms from the previously characterized LARP6 proteins of animals and vascular plants, thus establishing a distinct third class of the LARP6 protein family. 

Crustaceans are particularly sensitive to copper toxicity, and although the downstream effects of increased copper exposure on the metabolome are often postulated and observed, they are rarely measured. To perform absolute quantification of hydrophilic small-molecule metabolites in the hemolymph of the crustacean Cancer borealis, we derivatized targeted metabolites related to copper toxicity using in-house-developed isotopic N,N-dimethyl leucine (iDiLeu) tags. Selected analytes were pooled at previously determined concentrations to serve as internal standards, and a calibration curve was generated. The sample loss was minimized by optimizing the derivatization-assisted sample cleanup using dispersive liquid–liquid microextraction (DLLME) and hydrophilic–lipophilic balancing (HLB). Calibration curves were then used for the absolute quantification of metabolites of interest following 30 min, 1 h, and 2 h exposures to 10 µM CuCl2. We found that glutamic acid was downregulated after 2 h of copper exposure, which may disrupt cellular metabolism and increase oxidative stress in crustaceans. These changes could have significant impacts on crustacean populations and the ecosystems they support. 

ABSTRACT Multidrug-resistant (MDR) bacteria pose a significant public health challenge, underscoring the urgent need for innovative antibacterial strategies. Bacteriophages (phages), viruses that specifically target bacteria, offer a promising alternative; however, bacterial immune defenses often limit their effectiveness. Developing small-molecule inhibitors of these defenses can facilitate mechanistic studies and serve as adjuvants to enhance phage therapy. Here, we identify novel inhibitors targeting the bacterial cyclic oligonucleotide-based anti-phage signaling system (CBASS) effector Cap5. Cap5 is an HNH endonuclease activated by a cyclic nucleotide to degrade genomic DNA in virally infected cells, leading to cell death through abortive infection. Guided by the crystal structure of the Cap5 SAVED domain bound to its activating ligand, we performed structure-guided virtual screening to identify candidate inhibitors. Biochemical assays revealed that approximately 16% of the top docking hits exhibited inhibitory activity. Further cellular assays demonstrated that one potent compound could enterE. colicells and inhibit Cap5 activity. Our integrated approach—combining structure-based virtual screening with biochemical validation—provides a robust framework for discovering small-molecule inhibitors of bacterial immune defenses to advance adjunctive therapies and deepen our understanding of phage-bacteria interactions. 

Condensed matter studies have shown that fusion of two lipid membranes requires drastic structural rearrangements and is thus intrinsically slow. Interestingly, all forms of life on Earth use fusion to carry out some of the most fundamental life processes—communication, growth, metabolic homeostasis—that must be accomplished on timescales as short as a fraction of a millisecond. How do living systems beat the prohibitively slow speed limit imposed by membrane fusion? How do they tune the fusion timescale so that it matches a particular biological function? Here it is argued that fusion-mediated life processes as diverse as viral infection, muscle growth, and neuronal communication have all evolved at a common strategy that can be captured through a unifying relationship between the timescale of the process and the strength of the relevant trigger. Activated motion in a bias field along an emergent collective coordinate provides a suitable physical picture. The timescale is set by a reduced quantity defined as the trigger strength in the active state scaled by a system-specific critical parameter. The unified description suggests simple physical principles that organize the complexity of living systems and evolutionarily drive them toward functional behavior. 

Major and trace element geochemistry of lower Pliocene marine sediment core samples collected at International Ocean Discovery Program Site U1533 in the Amundsen Sea via ICP-MS. Bulk samples of mud or sandy mud were analyzed to assess sediment provenance using elemental ratios. The geochemical data were collected and analyzed by Olga Libman-Roshal and Sandra Passchier, assisted by Xiaona Li in the ICP-MS analytical work.