Transcription profiling by array of yeast strain BY4741 treated with alpha-factor mating pheromone to investigate nucleosome plasticity in G1 stage
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ABSTRACT:
ORGANISM(S): Saccharomyces cerevisiae
SUBMITTER: Oscar Flores
PROVIDER: E-MTAB-2195 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:Yeast strain BY4741 was grown overnight at 30C in YPD rich media. The yeast culture was diluted to an OD600 of 0.1 using fresh YPD media and further grown until an OD600 of 0.2. Then, alpha -factor mating pheromone (GenScript) was added to a final concentration of 10 uM to allow cell synchronization at G1 phase. After 2.5 h, the alpha-factor was removed by harvesting the cells for 10 min at 6000 rpm and decanting supernatant. The arrested cells were inoculated in fresh YPD rich medium to be released from G1-arrest. Samples were collected at 0, 30, 40, 50, and 70 mins, 7.5 ml samples were collected for RNA extraction while 40 ml samples were taken for nucleosomal DNA preparation and for flow cytometry (FACS). Samples for RNA isolation were collected by pipetting the culture directly into 15-ml Falcon tubes containing 7.5 ml of icy-water to quickly chill the cells. Cells were harvested by spinning for 3-4 min at 6000 rpm, frozen in liquid N2 and stored at - 80C. Total cellular RNA was extracted using the RNeasy kit (Qiagen), following the manufacturerM-^Rs instructions with the spheroplasting protocol. Purified RNA samples were quantified by Qubit fluorometer (Invitrogen, Inc.) and Nanodrop spectrophotometer (Thermo Scientific, Inc.). The total RNA was hybridized to Affymetrix GeneChip Yeast Genome 2.0 arrays for gene expression analysis.
2012-09-01 | E-MEXP-3702 | biostudies-arrayexpress
Project description:Fission yeast cells undergo sexual differentiation in response to nitrogen starvation. In this process haploid M and P cells first mate to form diploid zygotes, which subsequently enter meiosis and sporulate. Prior to mating, M cells secrete M-factor and P-cells secrete P-factor, and these diffusible mating pheromone can activate a signal transduction pathway in the opposite cell type. The pheromone response orchestrates mating and is also required for entry into meiosis. Here we use DNA microarrays to identify genes that are induced by M-factor in P cells and by P-factor in M-cells. The use of a cyr1 genetic background allowed us to study pheromone stimulation independently of nitrogen starvation. We identified a total of 163 genes that were consistently induced more than two-fold by pheromone stimulation. As expected, there was a substantial overlap between the genes induced by M- and P-factor. Surprisingly, we found that pheromone control extended to genes fulfilling their function well beyond the point of entry into meiosis, including numerous genes required for meiotic recombination. A direct comparison of the M- and P-factor induced expression pattern allowed us to identify cell-type specific transcripts, including three new M-specific genes and one new P-specific gene. Finally, our results support the notion that Ste11 is the key transcription factor activated by pheromone signalling. Keywords: dose response Gene expression profiling experiment in which gene expression in pheromone treated cultures was compared to that of untreated control cells. All cultures were growing exponentially in MSL minimal medium. S. pombe h+ cyr1- cells treated with M-factor mating pheromone for 2, 4 and 6 hrs respectively, were compared to non-treated cells. Similarly, S. pombe h- sax2- cyr1- cells treated with P-factor mating pheromone for 2, 4 and 6 hrs, were compared to non-treated cells. For identiofication of cell-type specific gene expression, h+ cyr1- cells were compaired to h+ sxa2- cyr1- cells. This was done both before pheromone treatment (0h/0h) and after 6 hours of pheromone treatment (6h/6h). RNA was extracted and subjected to cDNA expression profiling analysis using the established protocols (Xue et al, Yeast 21, 25-39, 2004).Data normalized with 'Lowess' per chip per spot normalization.
2007-08-21 | E-GEOD-5323 | biostudies-arrayexpress
Project description:Candida albicans can stochastically switch between two phenotypes, white and opaque. Opaque cells are the sexually competent form of C. albicans and therefore undergo efficient polarized growth and mating in the presence of pheromone. In contrast, white cells cannot mate, but are induced - under a specialized set of conditions - to form biofilms in response to pheromone. In this work, we compare the genetic regulation of such "pheromone-stimulated" biofilms with that of "conventional" C. albicans biofilms. In particular, we examined a network of six transcriptional regulators (Bcr1, Brg1, Efg1, Tec1, Ndt80, and Rob1) that mediate conventional biofilm formation for their potential roles in pheromone-stimulated biofilm formation. We show that four of the six transcription factors (Bcr1, Brg1, Rob1, and Tec1) promote formation of both conventional and pheromone-stimulated biofilms, indicating they play general roles in cell cohesion and biofilm development. In addition, we identify the master transcriptional regulator of pheromone-stimulated biofilms as C. albicans Cph1, ortholog of Saccharomyces cerevisiae Ste12. Cph1 regulates mating in C. albicans opaque cells, and here we show that Cph1 is also essential for pheromone-stimulated biofilm formation in white cells. In contrast, Cph1 is dispensable for the formation of conventional biofilms. The regulation of pheromone- stimulated biofilm formation was further investigated by transcriptional profiling and genetic analyses. These studies identified 206 genes that are induced by pheromone signaling during biofilm formation. One of these genes, HGC1, is shown to be required for both conventional and pheromone-stimulated biofilm formation. Taken together, these observations compare and contrast the regulation of conventional and pheromone-stimulated biofilm formation in C. albicans, and demonstrate that Cph1 is required for the latter, but not the former. 4 condition experiment: white and opaque cells in planktonic and pheromone-induced biofilm conditions with and without alpha pheromone. WT strain (P37005), the tec1 mutant strain and the cph1 mutant strain
2013-02-22 | E-GEOD-44449 | biostudies-arrayexpress
Project description:Expression profiling of honey bee brains exposed to queen mandibular pheromone. Exposure was performed in cages for 3 and 4 days; 3 and 4 days-old bees were analyzed
2011-09-01 | E-TABM-1102 | biostudies-arrayexpress
Project description:Modes of sexual reproduction in eukaryotic organisms are highly diversified. The human fungal pathogen Candida albicans undergoes a phenotypic switch from the white to the opaque phase in order to become mating-competent. In this study, we report that functionally and morphologically differentiated white and opaque cells show a coordinating behavior in the process of mating. Although white cells are mating-incompetent, they are induced to produce sexual pheromones when treated with opposite pheromones or interacted with opaque cells of an opposite mating type. In a co-culture system, pheromones released by white cells induce opaque cells to form mating projections and thus facilitate both opposite- and same-sex mating of opaque cells. Deletion of genes encoding the pheromone precursor proteins and inactivation of the pheromone response signaling pathway (Ste2-MAPK-Cph1) impair the promoting role of white cells (MTLa) in sexual mating of opaque cells. White and opaque cells communicate via a paracrine pheromone signaling and thus create an environment conducive to sexual mating. This coordination behavior of the two different cell types may be a trade-off strategy between sexual and asexual lifestyles in C. albicans. total RNA profiles of white cell treated with pheromone
2014-10-23 | E-GEOD-56039 | biostudies-arrayexpress
Project description:Disruption of STE18 gene which encodes a γ subunit of a heterotrimeric G protein in Candida albicans is shown to block pheromone-induced gene expression. Ectopic expression of either the Gα or the Gβ subunit of the heterotrimeric G protein is able to restore pheromone-induced gene expression. The transcriptional response to pheromone treatment was measured in the wild type strain, in a strain deleted for STE18, and in STE18 deleted strains ectopic expressing either the Gα or the Gβ subunit of G protein in opaque forms under RT.
2014-01-14 | E-GEOD-54031 | biostudies-arrayexpress
Project description:Comparison of female pheromone glands in Agrotis segetum and Agrotis ipsilon
2011-01-01 | E-MEXP-1419 | biostudies-arrayexpress
Project description:Transcriptional analysis of the pheromone gland in comparison to the rest of the insect body in female Agrotis segetum (Noctuidae).
2009-04-21 | E-MEXP-1418 | biostudies-arrayexpress
Project description:Expression profiling of honey bee brains exposed to brood pheromone. Exposure was performed in colonies and young (5 days-old) and old bees (15 days-old) were analyzed .
2008-12-31 | E-TABM-512 | biostudies-arrayexpress
Project description:Yeast cells were treated with different doses of pheromone in order to obtain dose-response profiles for induced genes. Specifically, dose-dependencies of transcriptional feedback regulators of the pheromone-response pathway were quantified.
2020-06-04 | GSE151729 | GEO