Comment[GEAAccession]	E-GEAD-475
MAGE-TAB Version	1.1
Investigation Title	Unveiling the intracellular stress responses, introduced and induced by higher level of glutathione using S. cerevisiae
Experiment Description	Glutathione (GSH) is a thiol and low-molecular-weight tripeptide consisted of Glu, Cys, and Gly. GSH is the most predominant anti-oxidant among various thiol molecules, and widely exists with milli molar level in prokaryotes and eukaryotes including mammalian. Importantly, the levels of GSH and ratio GSH/GSSG are steadily regulated to proceed or maintain many essential intracellular activities, i.e., GSH homeostasis. Many researches so far have proven that GSH-depleted cells can be responsible with aging, tumor, type-II diabetes, or neurodegenerative. While, as little research has been done that higher level of GSH could be loaded acutely onto cells (GSH stress), there is no clue how cells respond to and what factor would engage with GSH stress. In this study, we intend to partially unveil GSH stress-responsive mechanism using S. cerevisiae for model of mammalian cell. Here we performed RNA-seq analysis, and tried to screen pivotal factors dependent on GSH-stress response in yeast.
Experimental Design	compound treatment design	genetic modification design	genotyping design	normalization testing design	quality control testing design
Experimental Factor Name	genotype	compound	biological_replicate
Experimental Factor Type	genotype	compound	biological_replicate
Person Last Name	Yasukawa
Person First Name	Taishi
Person Affiliation	Mitsubishi Corporation Life Sciences Limited
Person Roles	submitter
Public Release Date	2023-11-06
Protocol Name	P-GEAD-1059	P-GEAD-1060	P-GEAD-1061	P-GEAD-1062	P-GEAD-1063
Protocol Type	sample collection protocol	nucleic acid extraction protocol	nucleic acid library construction protocol	nucleic acid sequencing protocol	normalization data transformation protocol
Protocol Description	Cultivated Sc cells in synthetic medium were harvested at log-phase by centrifugation, and the supernatant was removed. The cells were frozen immediately by liquid nitrogen, followed by the conservation in -80 degrees C.	Cells were treated with hot phenol method for conventional RNA purification method, and total RNA was purified using RNasey Mini kit (Qiagen).	Total RNA was quality-checked by Agilent 2100 Bioanalyzer and NanoDrop 2000. The mRNA libraries were constructed using NEBNext Ultra RNA Library Prep Kit for Illumina.	Illumina HiseqX sequencer was performed to read 151 base pair-ends method. Sequencing data was obtained until whose size had approached about 2 Gb per sample.	Toward Data, of which Q30 was verified to be more than 80%, some normalization steps were done using analytical tools following; featureCounts (BY4741_JRIS00000000.gff for reference), edgeR, perl, and R packages.
SDRF File	E-GEAD-475.sdrf.txt
Comment[AEExperimentType]	RNA-seq of coding RNA from single cells
Comment[BioProject]	PRJDB12811
Comment[Last Update Date]	2023-11-06