Project description:We have performed a systematic examination of genome-wide expression profiles using microarrays to reconstruct a global picture of the trx regulatory gene network in D. melanogaster. Using computational analysis of the microarray data, we have identified 25 clusters of genes potentially regulated by trx, most of them being located in the arm L of chromosome 3. Functional analysis revealed that most clusters are enriched in structural proteins involved in cuticle formation, which are preferently expressed in salivary glands. The same organization in clusters was observed in the transcriptomes of four independent experiments, being a distinctive feature of the regulatory networks of trx and other chromatin regulators (ASH2, NURF, Pc, ASH1). We have also identified many of these clusters in D. simulans, D. yakuba, D. pseudoobscura and partially in A. gambiae. Keywords: loss of function analysis
Project description:We have performed a systematic examination of genome-wide expression profiles using microarrays to reconstruct a global picture of the trx regulatory gene network in D. melanogaster. Using computational analysis of the microarray data, we have identified 25 clusters of genes potentially regulated by trx, most of them being located in the arm L of chromosome 3. Functional analysis revealed that most clusters are enriched in structural proteins involved in cuticle formation, which are preferently expressed in salivary glands. The same organization in clusters was observed in the transcriptomes of four independent experiments, being a distinctive feature of the regulatory networks of trx and other chromatin regulators (ASH2, NURF, Pc, ASH1). We have also identified many of these clusters in D. simulans, D. yakuba, D. pseudoobscura and partially in A. gambiae. Keywords: loss of function analysis
Project description:We have performed a systematic examination of genome-wide expression profiles using microarrays to reconstruct a global picture of the trx regulatory gene network in D. melanogaster. Using computational analysis of the microarray data, we have identified 25 clusters of genes potentially regulated by trx, most of them being located in the arm L of chromosome 3. Functional analysis revealed that most clusters are enriched in structural proteins involved in cuticle formation, which are preferently expressed in salivary glands. The same organization in clusters was observed in the transcriptomes of four independent experiments, being a distinctive feature of the regulatory networks of trx and other chromatin regulators (ASH2, NURF, Pc, ASH1). We have also identified many of these clusters in D. simulans, D. yakuba, D. pseudoobscura and partially in A. gambiae. Keywords: loss of function analysis Total RNA from w1118;+;+ larvae was pooled and used as a common reference in four microarrays against w1118;+;ash2I1 total RNA coming from two different extractions to take biological differences into account. Amplified RNA (aRNA) was obtained with the Amino-Allyl Messageamp II aRNA Amplification Kit (Ambion, Inc) from w1118;+;+ and w1118;+;ash2I1 prior to hybridization.
Project description:We have analyzed changes in histone H3 lysine 4 methylation (H3K4me) and gene expression resulting from trx knockdown (KD) in adult mushroom body neurons of Drosophila melanogaster.
Project description:We have performed a systematic examination of genome-wide expression profiles using microarrays to reconstruct a global picture of the trx regulatory gene network in D. melanogaster. Using computational analysis of the microarray data, we have identified 25 clusters of genes potentially regulated by trx, most of them being located in the arm L of chromosome 3. Functional analysis revealed that most clusters are enriched in structural proteins involved in cuticle formation, which are preferently expressed in salivary glands. The same organization in clusters was observed in the transcriptomes of four independent experiments, being a distinctive feature of the regulatory networks of trx and other chromatin regulators (ASH2, NURF, Pc, ASH1). We have also identified many of these clusters in D. simulans, D. yakuba, D. pseudoobscura and partially in A. gambiae. Keywords: loss of function analysis Total RNA from w1118;+;+ larvae was pooled and used as a common reference in four microarrays against w1118;+;trxE3/trxB11 total RNA coming from two different extractions to take biological differences into account. Two amplifications from w1118;+;+ and one from each w1118;+;trxE3/trxB11 replicate were performed with the Amino-Allyl Messageamp II aRNA Amplification Kit (Ambion, Inc) to obtain amplified RNA (aRNA). The two arrays from each replicate pair were hybridized with the same amplified RNA from sample and common reference but with dyes (Cy3 and Cy5 from Amersham, Inc) swapped to take dye-bias into account.
Project description:We have analyzed changes in histone H3 lysine 4 methylation (H3K4me) and gene expression resulting from trx knockdown (KD) in adult mushroom body neurons of Drosophila melanogaster.
Project description:We have analyzed changes in histone H3 lysine 4 methylation (H3K4me) and gene expression resulting from trx knockdown (KD) in adult mushroom body neurons of Drosophila melanogaster.
Project description:Pv11 is an insect cell line established from Polypedilum vanderplanki with ability to survive complete desiccation if they were treated with trehalose. Previous studies showed contribution of HSF1 gene in this adaptation. To determine HSF1 regulatory network in Pv11 cells we prepared RNAseq libraries for wild type Pv11 (WT), HSF1 knoked out (Hsf1-/-), HSf1 resqued (Hsf1-/-; Hsf1-HaloTag/BlaR), and negative control for HSF1 resqued cells (Hsf1-/- HaloTag/BlaR) under normal conditions and under incubation with trehalose for 48 h. Comparison of mRNA profiles showed that HSF1 induced anhydrobiosis-related genes, especially LEA and TRX, but suppressed cellular process-related genes, for example transcription- and translation-related genes.
Project description:The Caenorhabditis elegans oxidative stress response transcription factor, SKN-1, is essential for the maintenance of redox homeostasis and is a functional ortholog of the Nrf family of transcription factors. The numerous levels of regulation that govern these transcription factors underscore their importance. Here, we add a thioredoxin, encoded by trx-1, to the expansive list of SKN-1 regulators. We report that loss of trx-1 promotes nuclear localization of intestinal SKN-1 in a redox-independent, cell non-autonomous fashion from the ASJ neurons. Furthermore, this regulation is not general to the thioredoxin family, as two other C. elegans thioredoxins TRX-2 and TRX-3 do not play a role in this process. Moreover, TRX-1-dependent regulation requires signaling from the p38 MAPK signaling pathway. However, while TRX-1 regulates SKN-1 nuclear localization, SKN-1 transcriptional activity remains largely unaffected. Interestingly, RNA-Seq revealed that loss of trx-1 elicits a general, organism-wide down-regulation of several classes of genes; those encoding for collagens and lipid transport and localization being most prevalent. However, one prominent lipase-related gene, lips-6, is highly up regulated upon loss of trx-1 in a skn-1-dependent manner. Together, these results uncover a novel role for a thioredoxin in regulating intestinal SKN-1 nuclear localization in a cell non-autonomous manner, thereby contributing to the understanding of the processes involved in maintaining redox homeostasis throughout an organism. Four samples were analyzed: Two nematode strains were analyzed, each under non-stressed and stressed (10mM NaAs) conditions