Project description:Convergent microbial biocatalysis has emerged as a promising approach for the conversion of lignin side-streams into value-added chemicals in recent decades. However, the current knowledge of metabolic pathways directing the bioconversion of lignin-related aromatics is still limited to a few microbial species and unavailable for some of these compounds. Thus, the aim of this study was to identify the genes involved in the bioconversion of aromatic compounds in Xanthomonas citri subsp. citri 306 (X. citri 306), a bacterium belonging to a compelling yet untapped genus for studies on lignin-related aromatics metabolism. For this purpose, we used an integrative approach including genome data mining, RNA-seq, enzymology and gene knockout studies. The RNA-seq analysis revealed a total of 278 to 1464 differentially expressed genes (DEGs) in the aromatic-containing conditions compared to the control XVM2m-glucose, evidencing the importance of these compounds in modulating various physiological processes of X. citri 306 beyond the pathways related to their metabolism. Moreover, this work revealed the operon molRKAB, which plays a role in the first catabolic steps of the three main monolignols (p-coumaryl, coniferyl and sinapyl alcohols), besides showing all the enzymatic steps funneling them up to the tricarboxylic acid cycle. Additionally, the study uncovered aryl aldehyde reductases and efflux strategies that likely function to protect the pathogen from aromatics toxicity. Together, these findings enhance the current understanding of Xanthomonas metabolism and transcriptional responses to lignin-related aromatic compounds, shedding light on the diverse metabolic pathways available to enable the engineering of microbial chassis dedicated to lignin valorization.
Project description:In this study, we showed that three bacteria were able to inhibit the mycelial growth of the phytopathogenic fungus Thielaviopsis ethacetica, by the emission of microbial volatile organic compounds (mVOCs). Aiming to understand the molecular mechanisms of these interactions, we evaluated the transcriptomic response of T. ethacetica to the mVOCs produced by one of these bacterial isolates.
Project description:Identification of newer compounds to modulate dendritic cell functions. Total RNA obtained from bone marrow-derived dendritic cells treated for 6 hours with small chemical compounds or vehicle alone in the presence or absence of lipopolysaccharide (LPS).
Project description:Myocardin-related transcription factors (MRTFs) are coactivators of serum response factor (SRF), and thereby regulate cytoskeletal gene expression in response to actin dynamics. MRTFs have also been implicated in heat shock protein (hsp) transcription in fly ovaries, but the mechanisms remain unclear. Here we demonstrate that in mammalian cells, MRTFs are dispensable for hsp gene induction. However, the widely used small molecule inhibitors of MRTF/SRF transcription pathway, derived from CCG-1423, efficiently inhibit hsp gene transcription in both fly and mammalian cells also in absence of MRTFs. Quantifying RNA synthesis and RNA polymerase distribution demonstrates that CCG-1423-derived compounds have a genome-wide effect on transcription. Indeed, tracking nascent transcription at nucleotide resolution reveals that CCG-1423-derived compounds reduce RNA polymerase II elongation, and severely dampen the transcriptional response to heat shock. The effects of CCG-1423-derived compounds therefore extend beyond the MRTF/SRF pathway into nascent transcription, opening novel opportunities for their use in transcription research.
Project description:Myocardin-related transcription factors (MRTFs) are coactivators of serum response factor (SRF), and thereby regulate cytoskeletal gene expression in response to actin dynamics. MRTFs have also been implicated in heat shock protein (hsp) transcription in fly ovaries, but the mechanisms remain unclear. Here we demonstrate that in mammalian cells, MRTFs are dispensable for hsp gene induction. However, the widely used small molecule inhibitors of MRTF/SRF transcription pathway, derived from CCG-1423, efficiently inhibit hsp gene transcription in both fly and mammalian cells also in absence of MRTFs. Quantifying RNA synthesis and RNA polymerase distribution demonstrates that CCG-1423-derived compounds have a genome-wide effect on transcription. Indeed, tracking nascent transcription at nucleotide resolution reveals that CCG-1423-derived compounds reduce RNA polymerase II elongation, and severely dampen the transcriptional response to heat shock. The effects of CCG-1423-derived compounds therefore extend beyond the MRTF/SRF pathway into nascent transcription, opening novel opportunities for their use in transcription research.
Project description:High-content screening identified compounds that promote maturation of human neurons derived from pluripotent stem cells. We sequenced total RNA from neurons treated with these drugs to detemrine their effect at the transcriptome level.
Project description:Modulating beta-catenin signaling has attractive therapeutic potential in cancer immunotherapy. Various studies have found that beta-catenin can mediate immune evasion in cancer and promote anti-inflammatory features of antigen-presenting dendritic cells. Multiple small-molecule compounds that inhibit Wnt/beta-catenin signaling are currently in clinical development, but none have reached routine clinical use. Thus, new inhibitors of beta-catenin signaling are desirable. This dataset is the result of an investigation of the effect of small molecular compounds axitinib, ICG-001, nitazoxanide, orlistat, and XAV-939 on the maturation capacity of monocyte-derived dendritic cells (moDC). Treatment of immature moDC was done in combination with the beta-catenin activator 6-BIO. Immature moDCs were obtained by isolating monocytes from PBMC using magnetic beads, which were then differentiated into moDCs using GM-CSF and IL-4 for four days. Compounds were added during the maturation of the moDCs, which was done by adding LPS to the cell culture. After the maturation period of 24 hours, the mature moDCs were collected and processed for RNA sequencing.