Dectin-1, the major β-glucan receptor in leukocytes, triggers an effective immune response upon fungal recognition. Here we use sortase-mediated transpeptidation, a technique that allows placement of variety probes on polypeptide backbone, to monitor behavior labeled functional dectin-1 live cells with and without challenge. Installation by sortagging permitted highly specific visualization protein cell surface its subsequent internalization ligand presentation. Retrieval sortagged expressed...

Short-Chain Fatty Acids (SCFAs) are products of microbial fermentation that important for intestinal epithelial health. Here we describe SCFAs have rapid and reversible effects on Toll-like receptor (TLR) responses in cells. Incubation HEK293 or HeLa cells with the butyrate propionate at physiological concentrations enhanced NF-κB activation induced by TLR5, TLR2/1, TLR4 TLR9 agonists. response to TNFα was also increased SCFAs. Comparative transcript analysis HT-29 colon revealed...

Mucins play an essential role in protecting the respiratory tract against microbial infections while also acting as binding sites for bacterial and viral adhesins. The heavily O-glycosylated gel-forming mucins MUC5AC MUC5B eliminate pathogens by mucociliary clearance. Transmembrane MUC1, MUC4, MUC16 can restrict invasion at apical surface of epithelium. In this study, we determined impact host mucin glycans on epithelial entry SARS-CoV-2. Human lung Calu-3 cells express SARS-CoV-2 receptor...

Mucin 1 (MUC1) is a transmembrane mucin expressed at the apical surface of epithelial cells mucosal surfaces. MUC1 has barrier function against bacterial invasion and well known for its aberrant expression glycosylation in adenocarcinomas. The extracellular domain contains variable number tandem repeats (VNTR) 20 amino acids, which are heavily O -linked glycosylated. Monoclonal antibodies VNTR powerful research tools with applications diagnosis treatment MUC1-expressing cancers. Here, we...

Phagocytosis of the opportunistic fungal pathogen Candida albicans by cells innate immune system is vital to prevent infection. Dectin-1 major phagocytic receptor involved in anti-fungal immunity. We identify two new interacting proteins macrophages, Bruton's Tyrosine Kinase (BTK) and Vav1. BTK Vav1 are recruited cups containing C. yeasts or hyphae but absent from mature phagosomes. localize cuff regions surrounding hyphae, while lines full length phagosome. colocalize with lipid PI(3,4,5)P3...

The ability of phagocytes to clear pathogens is an essential attribute the innate immune response. role signaling lipid molecules such as phosphoinositides well established, but membrane sphingolipids in phagocytosis largely unknown. Using a genetic approach and small molecule inhibitors, we show that Candida albicans requires intact sphingolipid biosynthetic pathway. Blockade serine-palmitoyltransferase (SPT) ceramide synthase-enzymes involved biosynthesis- by myriocin fumonisin B1,...

Dysbiosis of the vaginal microbiome as a result overgrowth anaerobic bacteria leads to bacterial vaginosis (BV) which is associated with increased inflammation in genital mucosa. Moreover, BV increases susceptibility sexual transmitted infections (STIs) and adverse pregnancy outcomes. It remains unclear how specific aerobic affect health disease. We selected different ranging from true commensals species dysbiosis investigated their effects on activation dendritic cells (DCs). Commensal...

The cellular invasion machinery of the enteric pathogen Salmonella consists a type III secretion system (T3SS) with injectable virulence factors that induce uptake by macropinocytosis. at apical surface intestinal epithelial cells is inefficient, presumably because glycosylated barrier formed transmembrane mucins prevents T3SS contact host cells. We observed capable express mucin MUC1. Knockout MUC1 in HT29-MTX or removal sialic acids neuraminidase treatment reduced but did not affect...

Glycosylated mucin proteins contribute to the essential barrier function of intestinal epithelium. The transmembrane MUC13 is an abundant glycoprotein with important functions for mucosal maintenance that are not yet completely understood. We demonstrate in human epithelial monolayers, localized both apical surface and tight junction (TJ) region on lateral membrane. deletion resulted increased transepithelial resistance (TEER) reduced translocation small solutes. TEER buildup ΔMUC13 cells...

Sortagging is a versatile method for site-specific modification of proteins as applied to variety in vitro reactions. Here, we explore possibilities adapting the sortase use living cells. For intracellular sortagging, employ Ca²⁺-independent A transpeptidase (SrtA) from Streptococcus pyogenes. Substrate were equipped with C-terminal sortase-recognition motif (LPXTG); used an N-terminal (oligo)glycine nucleophiles. We show that sortase-dependent protein ligation can be achieved Saccharomyces...

ABSTRACT Bacterial vaginosis (BV) is a polymicrobial infection of the female reproductive tract. BV characterized by replacement health-associated Lactobacillus species diverse anerobic bacteria, including well-known Gardnerella vaginalis. Prevotella timonensis, and bivia are anerobes that found in significant number patients, but their contributions to disease process remain be determined. Defining characteristics overgrowth adherence mucosal surface increased activity mucin-degrading...

Abstract Bacterial vaginosis (BV) is a polymicrobial infection of the female reproductive tract (FRT). BV characterized by replacement health-associated Lactobacillus species diverse anaerobic bacteria, including well-known Gardnerella vaginalis . Prevotella timonensis and bivia are anaerobes that found in significant percentage patients, but their pathogenic properties yet to be determined. Defining characteristics overgrowth adherence mucosal surface increased activity mucin-degrading...

ABSTRACT Young females are at high risk of acquiring HIV‐1 infections and an imbalance in the vaginal microbiome enhances susceptibility to infection. More insights into underlying mechanisms could open up new strategies prevent acquisition dissemination. Here, we investigated effect anaerobic bacteria associated with bacterial vaginosis (BV) on transmission by two distinct dendritic cell (DC) subsets, that is, inflammatory monocyte‐derived DCs (moDCs) primary CD1c + DCs. Notably, contrast...

Abstract Probiotic bacteria including Bifidobacterial species have the capacity to improve intestinal health, but underlying molecular mechanisms are often not understood. Bifidobacteria considered keystone a relatively low abundance in adult tract. Bifidobacterium colonization depends on degradation of host-derived carbohydrates, human milk oligosaccharides and mucin-associated oligosaccharides. Specific strains can enhance barrier integrity symptoms gastrointestinal disorders. We...

The glyoxylate cycle, a metabolic pathway required for generating C4 units from C2 compounds, is an important factor in virulence, both animal and plant pathogens. Here, we report the localization of key enzymes this isocitrate lyase (Icl1; EC 4.1.3.1) malate synthase (Mls1; 2.3.3.9), human fungal pathogen Candida albicans. Immunocytochemistry combination with subcellular fractionation showed that Icl1 Mls1 are localized to peroxisomes, independent carbon source used. Although lack consensus...

The major fungal pathogen of humans, Candida albicans, is exposed to reactive nitrogen and oxygen species following phagocytosis by host immune cells. In response these toxins, this fungus activates potent anti-stress responses that include scavenging nitrosative oxidative via the glutathione system. Here we examine differential roles two recycling enzymes in redox homeostasis, stress adaptation virulence C. albicans: reductase (Glr1) S-nitrosoglutathione (GSNOR), Fdh3. We show...

In eukaryotes, acetyl coenzyme A (acetyl-CoA) produced during peroxisomal fatty acid beta-oxidation needs to be transported mitochondria for further metabolism. Two parallel pathways acetyl-CoA transport have been identified in Saccharomyces cerevisiae; one is dependent on citrate synthase (Cit), while the other requires and mitochondrial carnitine acetyltransferase (Cat) activities. Here we show that human fungal pathogen Candida albicans lacks Cit, relying exclusively Cat activity of...

The pentose phosphate pathway (PPP) is the main source of NADPH in cell and therefore essential for maintenance redox balance anabolic reactions. produced by two dehydrogenases oxidative branch PPP: glucose-6-phosphate dehydrogenase (Zwf1) 6-phosphogluconate (Gnd1). We observed that commensal fungus Candida albicans these enzymes contain putative peroxisomal targeting signals (PTSs): Zwf1 has a PTS1, while annotated intron GND1 encodes PTS2. By subcellular fractionation fluorescence...

Carnitine is an essential metabolite that enables intracellular transport of fatty acids and acetyl units. Here we show the yeast Candida albicans can synthesize carnitine de novo, identify 4 genes pathway. Null mutants orf19.4316 (trimethyllysine dioxygenase), orf19.6306 (trimethylaminobutyraldehyde dehydrogenase), orf19.7131 (butyrobetaine dioxygenase) lacked their respective enzymatic activities were unable to utilize acids, acetate, or ethanol as a sole carbon source, in accordance with...