Project description:Purpose: Evaluate the transcriptional profile of genes involved in fungus-host interactions using RNA sequencing Methods: T. rubrum and HaCat cells line were co-cultured in RPMI medium supplemented with 5% of fetal bovine serum and were incubated for 24 h at 37ºC in a humidified atmosphere containing 5% CO2 , followed RNA by extraction of both organisms, libraries construction and sequence. Results: Our data demonstrated the induction of specific genes that may improve the assimilation of nutrients and fungal survival in the host and genes encoding keratinolytic proteases that are important for T. rubrum virulence during infection. In HaCat cell lines it were induced genes encoding antimicrobial activity , cell migration and epithelial barrier repair. Futhermore, the genes KRT1 and FLG involved in epithelial barrier integrity were repressed Conclusions: This mixed transcriptome analysis showed the modulation of important genes involved in the mechanism of T. rubrum infection and in the defense and maintenance of cell homeostasis of keratinocytes, which could represent potential antifungal targets for new therapeutic approaches to the treatment of dermatophytoses
Project description:We complemented our genomic investigations with keratinolytic activity and cellular metabolism reconsctruction, yielding a more comprehensive view of the degradation process.
Project description:Experience-dependent gene transcription is required for nervous system development and function. However, the DNA regulatory elements that control this program of gene expression are not well defined. Here we characterize the enhancers that function across the genome to mediate activity-dependent transcription in neurons. While ~12,000 putative activity-regulated enhancer sequences have previously been identified that are enriched for H3K4me1 and the histone acetyltransferase CBP, we find that this chromatin signature is not sufficient to distinguish which of these regulatory sequences are actively engaged in promoting activity-dependent transcription. We show here that a subset of H3K4me1/CBP positive enhancers that is enriched for H3K27 acetylation (H3K27ac) in vivo, and shows increased H3K27ac upon membrane depolarization of cortical neurons, function to regulate activity-dependent transcription. The function of many of these activity-regulated enhancers appears to be dependent on the binding of FOS, a protein that had previously been thought to interact primarily with the promoters of activity-regulated genes. Furthermore, many of these target genes in cortical neurons encode neuron specific proteins that regulate synaptic development and function. These findings suggest that FOS functions at enhancers to control activity-dependent gene programs that are critical for nervous system function, and provide a resource of activity-dependent enhancers that may give insight into genetic variation that contributes to brain development and disease. Genome-wide maps of H3K27ac and AP1 transcription factors (CFOS, FOSB, JUNB) before and after neuronal activity in mouse cortical neurons.