Temperature induced differentiation of piA immortalised cell line PrE2.8
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ABSTRACT: The cell line PrE2.8 was cultured at a permissive growth temperature of 33oC and then half transferred to a non-permissive temperature of 39.5oC for 4days. The two sets of cells were harvested and co-hybridised to identify genes with altered expression resulting from unduced differentiation. Keywords: Induction of cellular differentiation
Project description:We performed global scale microarray analysis to identify detailed mechanisms by which nonpermissive temperature induces cell growth arrest and differentiation in tracheal epithelial RTEC11 cells harboring temperature-sensitive simian virus 40 large T-antigen by using an Affymetrix GeneChip system. Tracheal epithelial RTEC11 cells used in this study were derived from transgenic rats harboring a temperature-sensitive simian virus 40 large T-antigen. Although the cells grew continuously at the permissive temperature, the nonpermissive temperature led to cell growth arrest and differentiation. Keywords: rat tracheal epithelial RTEC11; growth arrest; differentiation; gene expression
Project description:We performed global scale microarray analysis to identify detailed mechanisms by which nonpermissive temperature induces cell growth arrest and differentiation in astrocyte RCG-12 cells harboring temperature-sensitive simian virus 40 large T-antigen by using an Affymetrix GeneChip system. Astrocyte RCG-12 cells used in this study were derived from primary cultured rat cortical glia cells infecting with a temperature-sensitive simian virus 40 large T-antigen. Although the cells grew continuously at the permissive temperature, the nonpermissive temperature led to cell growth arrest and differentiation. Of the 15,923 probe sets analyzed, nonpermissive temperature differentially expressed 556 probe sets by >2.0-fold. Keywords: Astrocyte cell; growth arrest; differentiation; temperature-sensitive simian virus 40 large T-antigen; nonpermissive temperature
Project description:Expression analysis of Saccharomyces cerevisiae TAF5 and taf5 temperature conditional mutants grown at permissive and non-permissive temperature. Investigation of whole genome gene expression level changes in Saccharomyces cerevisae taf5-17, taf5-45, taf5-408 and taf5-10.4 mutants, compared to the wild-type strain. The mutations engineered into the strains confer temperature conditional growth. The mutants analyzed in this study are further described in Layer et. al., 2010. Direct Transactivator-Transcription Factor IID (TFIID) Contacts Drive Yeast Ribosomal Protein Gene Transcription. Journal of Biological Chemistry.
Project description:Measles virus infects serum activated airway epithelial cells and many adenocarcinoma cell lines. A microarray analysis was performed on virus permissive versus non-permissive cells. Membrane protein genes that were upregulated in permissive cells were tested as receptor/entry factors. Membrane protein genes that were upregulated in smooth airway epithelial cells (SAEC) following growth in 10% fetal calf serum that made the cell line permissive to measles virus were identified. Membrane protein genes that were upregulated in adenocarcinoma cells that were permissive to wild type measles virus infection were identified. [SAEC]: Airway cells (SAEC) grown in serum free media (SAGM) were purchaced from Lonza. Half the cells were cultured in SAGM, the other half were transferred into Dulbecco's 10% fetal calf serum for 24 hrs. RNA was harvested from the cells by the Qiagen RNAeasy [Adenocarcinoma cells]: MCF7, MDA-MB-468, T47D, NCI-H358, NCI-H125, MGH24 cells were permissive and A549 and MDA-MB-231 cells were non-permissive.
Project description:Wheat plants (T. aestivum cv. Chinese Spring) were grown in a greenhouse and watered daily to avoid drought stress. The main stem ears were labelled when flowering. At 12 days after anthesis, the plants were transferred to growth chambers with a temperature regime of 24/17◦C (14 h/10 h) day/night cycle. At 15 days after anthesis, half of the plants were applied a temperature regime of 37/17◦C for 3 days and were harvested on the 4th day after 4 h under heat with light; the remaining plants were kept at 24/17◦C as a control and were harvested at the same sampling time.
Project description:This is a study to characterize gene expression profiles in stored Russet Burbank potato tubers. Tubers were harvested from commercial fields in the central sands region of Wisconsin in the fall of 2006. The tubers were put into storage at 55 degrees F for preconditioning and wound healing. Shortly after the temperature of the storage bin began to decrease, uniform, healthy tubers were selected for use in this microarray analysis. Tubers were at 53.6 degrees F at this time, and pieces of starch-storing tissue were collected for use as the reference sample. Other tubers were moved to temperature-controlled lockers and these were cooled gradually to either 48 or 40 degrees F following industry standard procedures. The expectation was that tubers held at 48 degrees would not have a significant accumulation of glucose and fructose, but that tubers cooled to 40 degrees would undergo low temperature sweetening and accumulate glucose and fructose to a degree that is unsuitable for processing. Three weeks later, when the locker temperatures were 48 degrees F and 41.5 degrees F, tissue samples were collected for RNA analysis. After another three weeks, samples were collected from tubers at 48 degrees F and 40 degrees F. At that time some tubers were moved from the 48 degree locker to the 40 degree locker in order to see if gene expression changes observed as a result of gradual cooling are similar to those that occur following a sudden decrease in temperature. Three weeks later, samples were collected from tubers held at 48 degrees F, tubers held at 40 degrees F, and from the tubers that were moved from 48 to 40 degrees F. At this time another set of tubers was transferred from 48 degrees to 40 degrees. Three weeks later the last samples were harvested from tubers held at 48 degrees F, from tubers held at 48 degrees F and from tubers that were transferred three weeks prior from 48 to 40 degrees. RNA was isolated from tissue extracted from three tubers. Keywords: Reference design
Project description:Transcriptome analyses of thermosensitive exosome mutants from permissive temperature to various time exposure to restrictive temperature.
Project description:Expression analysis of Saccharomyces cerevisiae TAF5 and taf5 temperature conditional mutants grown at permissive and non-permissive temperature. Investigation of whole genome gene expression level changes in Saccharomyces cerevisae taf5-17, taf5-45, taf5-408 and taf5-10.4 mutants, compared to the wild-type strain. The mutations engineered into the strains confer temperature conditional growth. The mutants analyzed in this study are further described in Layer et. al., 2010. Direct Transactivator-Transcription Factor IID (TFIID) Contacts Drive Yeast Ribosomal Protein Gene Transcription. Journal of Biological Chemistry. A twenty chip study using total RNA recovered from four separate wild-type cultures of Saccharomyces cerevisiae and four separate cultures for each of four taf5 temperature conditional mutants (16 mutant cultures). Each chip measures the expression level of 5,777 genes from Saccharomyces S288C with eight 60-mer probe pairs (PM/MM) per gene, with three-fold technical redundancy.
Project description:To generate a bona fide model to study post-stress baterial programmed cell death (PCD), we used a temperature-sensitive E. coli mutant (dnaB-Ts) since temperature stress can be rapidly reversed while other stress, such as antibotic treatment, is difficult to be rapidly and completely stopped and removed for post-stress PCD analysis. When cells were shifted from permissive (30 °C) to non-permissive temperature (42 °C), the dnaB-Ts ΔahpC double mutant maintained full surival while survival of the dnaB-Ts single mutant dropped 3 orders of magnitude; similarly, the double mutant showed much higher survival during treatments with quinolones and β-lactams at permissive temperature. The purpose of the RNA-seq analyses is to uncover molecular mechanisms underlying the increased survival of the dnaB-Ts ΔahpC double mutant by comparing its transcriptional profiles with that of the dnaB-Ts single mutant at both permissive and non-permissive temperature. The results showed that multiple antioxidative systems and stress response pathways were highly upregulated due to a deficiency of ahpC when combined with a dnaB-Ts mutation. The high expression of these genes are consistent with the lower levels of toxic reactive oxygen species (ROS) in the dnaB-Ts ΔahpC mutant than in the dnaB-Ts single mutant. Thus, the RNA-seq data supported that ROS are an important lethal factor in bacterial suicide pathways when bacterial cells are exposed to harsh stress.
Project description:The target of rapamycin (TOR) plays a central role in eukaryotic cell growth control. With prevalent hyper-activation of the mTOR pathway in human cancers, novel strategies to enhance TOR pathway inhibition are highly desirable. We used a yeast-based high-throughput chemical genetic screen to identify small-molecule enhancers of rapamycin (SMERs) and used whole genome expression analysis to identify their mechanisms of action. We incubated Met30 temperature-sensitive yeast strain at either the permissive (room temperature) or non-permissive (35°C) temperature for 1 hour prior to RNA extraction and hybridization on Affymetrix microarrays. Comparison of expression profiles enabled identification of gene-signature characteristic of Met30 inhibition.