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Abstract Fungal plasma membrane proteins represent key therapeutic targets for antifungal agents, yet their native structure and spatial distribution remain poorly characterized. Herein, we employ an integrative approach to investigate the organization of plasma membrane protein complexes inCandida glabrata, focusing on two abundant and essential membrane proteins, the β-(1,3)-glucan synthase (GS) and the proton pump Pma1. We show that treatment with caspofungin, an echinocandin antifungal that targets GS, disrupts the native distribution of membrane protein complexes and alters membrane biophysical properties. Perturbation of the sphingolipid biosynthesis further modulates drug susceptibility, revealing that the lipid environment plays an integral role in membrane protein organization and GS-echinocandin interactions. Our work highlights the importance of characterizing membrane proteins in their native context to understand their functions and inform the development of novel antifungal therapies. 

This study uses cryo-electron microscopy to reveal the distinct nanoscale structures within protein condensates, highlighting the potential correlation between their internal organization and material properties. 

Precipitation changes altered soil heterotrophic respiration, but the underlying microbial mechanisms remain rarely studied. This study conducted three-year switchgrass (Panicum virgatum L.) mesocosm experiment to investigate soil heterotrophic respiratory responses to altered precipitation. Five treatments were considered, including ambient precipitation (P0), two wet treatments (P+33 and P+50: 33% and 50% enhancement relative to P0), and two drought treatments (P-33 and P-50: 33% and 50% reduction relative to P0). The plant’s aboveground biomass (AGB), soil organic carbon (SOC), total nitrogen (TN), microbial biomass carbon (MBC), heterotrophic respiration (Rs), biomass-specific respiration (Rss: respiration per unit of microbial biomass as a reciprocal index of microbial growth efficiency), and extracellular enzymes activities (EEAs) were quantified in soil samples (0–15 cm). Despite significantly different soil moisture contents among treatments, results showed no impact of precipitation treatments on SOC and TN. Increasing precipitation had no effect, but decreasing precipitation significantly reduced plant AGB. Relative to P0, P+33 significantly increased Rs by more than 3-fold and caused no changes in MBC, leading to significantly higher Rss (P < 0.05). P+33 also significantly increased hydrolytic enzyme activities associated with labile carbon acquisition (Cacq) by 115%. The only significant effect of drought treatments was the decreased β-D-cellobiosidase (CBH) and peroxidase (PEO) under P-33. Nonparametric analyses corroborated the strong influences of moisture and CBH on the enhanced precipitation, which stimulated soil respiratory carbon loss, likely driven by both elevated hydrolase activities and reduced microbial growth efficiency. However, the less sensitive drought effects suggested potential microbial tolerance to water deficiency despite depressed plant growth. This study informs the likely decoupled impacts of microbes and plants on soil heterotrophic respiration under changing precipitation in the switchgrass mesocosm experiment. 

ABSTRACT Dihydrouridine is an abundant and conserved modified nucleoside present on tRNA, but characterization and functional studies of modification sites and associated DUS writer enzymes in mammals is lacking. Here we use a chemical probing strategy, RNABPP-PS, to identify 5-chlorouridine as an activity-based probe for human DUS enzymes. We map D modifications using RNA-protein crosslinking and chemical transformation and mutational profiling to reveal D modification sites on human tRNAs. Further, we knock out individual DUS genes in two human cell lines to investigate regulation of tRNA expression levels and codon-specific translation. We show that whereas D modifications are present across most tRNA species, loss of D only perturbs the translational function of a subset of tRNAs in a cell type-specific manner. Our work provides powerful chemical strategies for investigating D and DUS enzymes in diverse biological systems and provides insight into the role of a ubiquitous tRNA modification in translational regulation.