Project description:The anti-mycobacterial activity of C17 diynes has been described previously, however, their mode of action remains unknown. Microarray techniques were used to explore the genetic regulation reponses of Mycobacterium smegmatis to treatment with the C17 diynes, falcarinol and panaxydol. Our analyses showed a distinct mode of action of the C17 diynes when compared with commonly used anti-mycobacterial drugs. In addition, geneset enrichment analysis, pathway enrichment analysis and PASS analysis revealed significant gene ontology terms, pathways and potential modes of action, respectively. Combing the results of the three analyses, we hypothesize that the C17 diynes inhibit fatty acid biosynthesis, specifically phospholipid synthesis in mycobacteira.
Project description:The anti-mycobacterial activity of C17 diynes has been described previously, however, their mode of action remains unknown. Microarray techniques were used to explore the genetic regulation reponses of Mycobacterium smegmatis to treatment with the C17 diynes, falcarinol and panaxydol. Our analyses showed a distinct mode of action of the C17 diynes when compared with commonly used anti-mycobacterial drugs. In addition, geneset enrichment analysis, pathway enrichment analysis and PASS analysis revealed significant gene ontology terms, pathways and potential modes of action, respectively. Combing the results of the three analyses, we hypothesize that the C17 diynes inhibit fatty acid biosynthesis, specifically phospholipid synthesis in mycobacteira. Mycobacterium smegmatis MC2 155 was treated with 10 times of MIC90 of falcarinol, panaxydol, isoniazid, ethambutol and kanamycin for 6 hours with at least 6 independent biological replicates.
Project description:The aim of this experiment was to investigate the gene expression profile of two xenografted nasopharyngeal carcinoma tumors - C15 and C17 which are international reference tools for biological investigations of this type of disease. C15 and C17 transcriptome was profiled by comparison with human untransformed diploid fibroblasts, MRC5. These fibroblasts are very sensitive to contact inhibition and were collected after 3 days of confluence, therefore in a quiescent state. We acknowledge that malignant nasopharyngeal carcinoma cells and MRC5 cells do not belong to the same lineage (epithelial cells versus fibroblasts). However this combination was expected to highlight genes involved in proliferation of NPC cells. Competitive hybridisation of cDNA for each pair C15 and MRC5, C17 and MRC5 was performed on Agilent microarrays containing 22 000 60mer oligonucleotides related to 16 000 human transcripts. The analysis was focused mainly on genes overexpressed in both C15 and C17 by comparison with MRC5
Project description:Six- to eight-week old CD1 male mice were sleep-deprived for 24 hours by placing them in a small platform in a water tank. Their ileum samples were harvested every 4 hours on the next day (time point: ZT1,ZT5,ZT9,ZT13,ZT17,ZT21). The ileum samples were subjected to 16s rDNA sequencing. Control group (C1,C5,C9,C13,C17,C21, n=36). SD group (SD1,SD5,SD9,SD13,SD17,SD21, n=36).
Project description:The dual-specificity tyrosine phosphorylation-regulated kinase DYRK2 has emerged as a key regulator of cellular processes such as proteasome-mediated protein degradation. To gain further insights into its function, we took a chemical biology approach and developed C17, a potent small-molecule DYRK2 inhibitor, through multiple rounds of structure-based optimization guided by a number of co-crystallized structures. C17 displayed an effect on DYRK2 at a single-digit nanomolar IC50 and showed outstanding selectivity for the human kinome containing 467 other human kinases. Using C17 as a chemical probe, we further performed quantitative phosphoproteomic assays and identified several novel DYRK2 targets, including eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1) and stromal interaction molecule 1 (STIM1). DYRK2 phosphorylated 4E-BP1 at multiple sites, and the combined treatment of C17 with AKT and MEK inhibitors showed synergistic 4E-BP1 phosphorylation suppression. The phosphorylation of STIM1 by DYRK2 substantially increased the interaction of STIM1 with the ORAI1 channel, and C17 impeded the store-operated calcium entry process. Collectively, these studies further expand our understanding of DYRK2 and provide a valuable tool to further pinpoint its biological function.
Project description:The dual-specificity tyrosine phosphorylation-regulated kinase DYRK2 has emerged as a key regulator of cellular processes such as proteasome-mediated protein degradation. To gain further insights into its function, we took a chemical biology approach and developed C17, a potent small-molecule DYRK2 inhibitor, through multiple rounds of structure-based optimization guided by a number of co-crystallized structures. C17 displayed an effect on DYRK2 at a single-digit nanomolar IC50 and showed outstanding selectivity for the human kinome containing 467 other human kinases. Using C17 as a chemical probe, we further performed quantitative phosphoproteomic assays and identified several novel DYRK2 targets, including eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1) and stromal interaction molecule 1 (STIM1). DYRK2 phosphorylated 4E-BP1 at multiple sites, and the combined treatment of C17 with AKT and MEK inhibitors showed synergistic 4E-BP1 phosphorylation suppression. The phosphorylation of STIM1 by DYRK2 substantially increased the interaction of STIM1 with the ORAI1 channel, and C17 impeded the store-operated calcium entry process. Collectively, these studies further expand our understanding of DYRK2 and provide a valuable tool to further pinpoint its biological function.