Project description:Here we performed DNAse-seq experiments on samples of adherent cultures of mouse neural stem cells (NS5 cell line) under normal growth conditions and upon differentiation by expression of an inducible version of the proneural factor Mash1/Ascl1 (Ascl1-ERT2). This resulted in the generation of genome-wide maps of regions of chromatin accessibility in both conditions.
Project description:Among the flaviviral proteins, NS5 is the largest and most conserved. NS5 contains major enzymatic components of the viral replication complex. Disruption of the common key NS5-host protein-protein interactions critical for viral replication could aid in the development of broad-spectrum anti-flaviviral therapeutics. To this end, we investigated the JEV- and ZIKV-NS5 interactomes in human cells using GFP pull-downs with mass spectrometry analysis in a label-free fashion. A total of 138 cellular proteins interacting with NS5 from JEV, ZIKV, or both were identified as Protein classification analysis of identified cellular targets revealed the enrichment of RNA binding, processing and splicing including spliceosomal and spliceosome-associated proteins in both datasets. Comparison of our data with literature not only revealed several cellular NS5 interacting proteins shared among flaviviruses, but also identified proteins that have no known function in flavivirus biology such as RNA polymerase II-associated Paf1 complex, protein phosphatase 6, and s-adenosylmethionine synthetase. Our study generates the first landscape of the JEV and ZIKV NS5 interactome in human cells and identifies cellular proteins that are potentially targetable for broad-spectrum anti-flaviviral therapy.
Project description:Gene expression in NS5 murine neural stem cells was compared to that of astrocytes derived from NS5 cells via 3 days of treatment with foetal calf serum (FCS). Keywords: cell type comparison
Project description:Here we performed a ChIP-seq experiment on a sample of adherent cultures of mouse neural stem cells (NS5 cell line) under normal growth conditions. This resulted in the generation of a genome-wide map of Zeb1 binding to chromatin.
Project description:Here we performed a ChIP-seq experiment on a sample of adherent cultures of mouse neural stem cells (NS5 cell line) under normal growth conditions. This resulted in the generation of a genome-wide map of RBPJ binding to chromatin.
Project description:We use ChIP-seq on PAF1C member Leo1 to determine how PAF1C occupancy at interferon stimulated genes is impacted by dengue virus NS5 protein.
Project description:Here we performed a ChIP-seq experiment on a sample of adherent cultures of mouse neural stem cells (NS5 cell line) under normal growth conditions and upon short term activation (30 minutes) of an inducible version of the proneural factor Mash1/Ascl1 (Ascl1-ERT2). This resulted in the generation of a genome-wide map of Ascl1 binding to chromatin.
Project description:The ChIP-seq and RNA-seq data provide evidence that ZIKV-NS5 binds to the gene body of the key neuro-factors and then inhibits their transcription, thus disrupting the neurogenesis of human NPCs.
Project description:Hypusination is a unique post-translational modification of the eukaryotic translation factor 5A (eIF5A) that is essential for overcoming ribosome stalling at polyproline sequence stretches. The initial step of hypusination, the formation of deoxyhypusine, is catalyzed by deoxyhypusine synthase (DHS), however, the molecular details of the DHS-mediated reaction remained elusive. Recently, patient-derived variants of DHS and eIF5A have been linked to rare neurodevelopmental disorders. Here, we present the cryo-EM structure of the human eIF5A-DHS complex at 2.8Å resolution and a crystal structure of DHS trapped in the key reaction transition state. Furthermore, we show that disease-associated DHS variants influence the complex formation and hypusination efficiency. Hence, our work dissects the molecular details of the deoxyhypusine synthesis reaction and reveals how clinically-relevant mutations affect this crucial cellular process.