Transcription profiling of chicken bursal cells treated with stem cell antigen 2
Ontology highlight
ABSTRACT: The differentiation and proliferation of chicken B cells rely on the antigens on the cell surface of bursal stromal cells. Chicken stem cell antigen 2 (SCA2) is localized on the surface of bursal cortical and medullary epithelial cells. The signals through SCA2 may regulate the expressions critical for the B cell development. The altered expressions of bursal cells that were induced through SCA2-signaling were detailed via Affemetrix array experiments. Experiment Overall Design: Bursa of 3 week old line 0 chickens were collected and manually teased to harvest the bursal cells. The bursal cells were spread on the monolayers of SCA2Flag-DF1 cells or control Vector-DF1 cells in triplicate. After growth in the mixture of Liebovitzâs L-15 and McCoy 5A media (1:1) supplemented with 4% FBS, penicillin, and streptomycin for overnight at 37 ºC in 5% CO2, the bursal cells were recovered by washes (DF1 cells are adhesive) followed by gentle centrifugation at 1000Ãg for 10 min. Total RNA was isolated using the absolutely RNA microprep kit.
Project description:The differentiation and proliferation of chicken B cells rely on the antigens on the cell surface of bursal stromal cells. Chicken stem cell antigen 2 (SCA2) is localized on the surface of bursal cortical and medullary epithelial cells. The signals through SCA2 may regulate the expressions critical for the B cell development. The altered expressions of bursal cells that were induced through SCA2-signaling were detailed via Affemetrix array experiments.
Project description:These libraries represent total RNA from bursal tissue of 20 day old CB-inbred chicks and the ALV induced bursal cell line DT40 Cre1 cells. Keywords = chicken transcriptome SAGE Bursal cell DT40 Keywords: other
Project description:MarekM-bM-^@M-^Ys disease (MD) is an economically significant disease in chickens caused by the highly oncogenic MarekM-bM-^@M-^Ys disease virus (MDV). A major unanswered question is the mechanism of MDV-induced tumor formation. Meq, a bZIP transcription factor discovered in the 1990s, is critically involved in viral oncogenicity but only a few of its host target genes have been described impeding our understanding of MDV-induced tumorigenesis. Using ChIP-seq and microarray analysis, a high confidence list of Meq-binding sites in the chicken genome and a global transcriptome of Meq-responsive genes was generated. Meq binding sites were found to be enriched in the promoter regions of up-regulated genes, but not in those of down-regulated genes. ChIP-seq was also performed for c-Jun, a known heterodimeric partner of Meq. Close location of binding sites of Meq and c-Jun was noted, suggesting cooperativity between these two factors in modulating transcription. Pathway analysis indicated that Meq transcriptionally regulates many genes that are part of several signaling pathways include the ERK/MAPK, Jak-STAT, and ErbB pathways that are critical for oncogenesis and/or include signaling mediators involved in apoptosis. Meq activates oncogenic signaling cascades by transcriptionally activating major kinases in the ERK/MAPK pathway and simultaneously repressing phosphatases, as verified using inhibitors of MEK and ERK1/2 in a cell proliferation assay. This study provides significant insights into the mechanistic basis of Meq-dependent cell transformation. Transcript profiling of DF-1 (a chicken embryo fibroblast cell line) and Meq-DF-1 clone 5G (DF-1 stably expressing Meq driven by the CMV promoter) using Affymetrix chicken GeneChips
Project description:Marekâs disease (MD) is an economically significant disease in chickens caused by the highly oncogenic Marekâs disease virus (MDV). A major unanswered question is the mechanism of MDV-induced tumor formation. Meq, a bZIP transcription factor discovered in the 1990s, is critically involved in viral oncogenicity but only a few of its host target genes have been described impeding our understanding of MDV-induced tumorigenesis. Using ChIP-seq and microarray analysis, a high confidence list of Meq-binding sites in the chicken genome and a global transcriptome of Meq-responsive genes was generated. Meq binding sites were found to be enriched in the promoter regions of up-regulated genes, but not in those of down-regulated genes. ChIP-seq was also performed for c-Jun, a known heterodimeric partner of Meq. Close location of binding sites of Meq and c-Jun was noted, suggesting cooperativity between these two factors in modulating transcription. Pathway analysis indicated that Meq transcriptionally regulates many genes that are part of several signaling pathways include the ERK/MAPK, Jak-STAT, and ErbB pathways that are critical for oncogenesis and/or include signaling mediators involved in apoptosis. Meq activates oncogenic signaling cascades by transcriptionally activating major kinases in the ERK/MAPK pathway and simultaneously repressing phosphatases, as verified using inhibitors of MEK and ERK1/2 in a cell proliferation assay. This study provides significant insights into the mechanistic basis of Meq-dependent cell transformation. ChiP-Seq of Meq-DF-1 clone 5G (DF-1 stably expressing Meq driven by the CMV promoter) with Meq and Jun antibodies
Project description:The plasma membrane proteome resides at the interface between the extra- and intra-cellular environment and through its various roles in signal transduction, immune recognition, nutrient transport, and cell-cell and cell-matrix interactions plays a critical role in determining the fate of a cell. The objective of this study was to characterize the cell-membrane proteome of SKBR3/HER2+ breast cancer cells by using mass spectrometry detection, to advance the understanding of biological triggers that sustain aberrant cell survival, proliferation, and development of resistance to therapeutic drugs.
Project description:Marekâs disease (MD) is an economically significant disease in chickens caused by the highly oncogenic Marekâs disease virus (MDV). Understanding the genes and biological pathways that confer MD genetic resistance should lead towards the development of more disease resistant commercial poultry flocks or improved MD vaccines. MDV Meq, a bZIP transcription factor, is largely attributed for viral oncogenicity though only a few host target genes have been described, which has impeded our understanding of MDV-induced tumorigenesis. Given the importance of Meq in MDV-induced pathogenesis, we explored the role of Meq in genetic resistance to MDV. Using global transcriptome analysis to compare the host response between birds challenge with either wild type MDV or a recombinant lacking Meq, we identified a number of specific genes and pathways associated with either MD resistance. Integrating prior information from ChIP-seq, microarray analysis, and SNPs exhibiting allele-specific expression (ASE) in response to MDV infection from two inbred layer lines that differ greatly in MD genetic resistance, we were able to provide a evidence for 35 genes that SNPs within transcription factor binding sites can affect transcription factor binding and gene expression in an allele-specific manner. Transcript profiling of CEF's (a chicken embryo fibroblast cell line) from MD susceptible and resistant lines using Affymetrix chicken GeneChips
Project description:We used a chicken immune-targeted gene array to analyse the differences in gene expression in the bursa of Fabricius from genetically resistant and susceptible animals infected with Infectious Bursal Disease Virus (IBDV).
Project description:These libraries represent total RNA from bursal tissue of 20 day old CB-inbred chicks and the ALV induced bursal cell line DT40 Cre1 cells. Keywords = chicken transcriptome SAGE Bursal cell DT40
Project description:The plasma membrane proteome resides at the interface between the extra- and intra-cellular environment and through its various roles in signal transduction, immune recognition, nutrient transport, and cell-cell and cell-matrix interactions plays a critical role in determining the fate of a cell. The objective of this study was to characterize the cell-membrane proteome of SKBR3/HER2+ breast cancer cells by using mass spectrometry detection, to advance the understanding of biological triggers that sustain aberrant cell survival, proliferation, and development of resistance to therapeutic drugs.
Project description:The plasma membrane proteome resides at the interface between the extra- and intra-cellular environment and through its various roles in signal transduction, immune recognition, nutrient transport, and cell-cell and cell-matrix interactions plays a critical role in determining the fate of a cell. The objective of this study was to characterize the cell-membrane proteome of SKBR3/HER2+ breast cancer cells by using mass spectrometry detection, to advance the understanding of biological triggers that sustain aberrant cell survival, proliferation, and development of resistance to therapeutic drugs.