Screening Of Gene Related To Mono(2-ethylhexyl) Phthalate Sensitivity In Saccharomyces Cerevisiae Genome

Mono(2-ethylhexyl)phthalate(MEHP)is a recognized animal endocrine disruptors,and research of MEHP mainly focus on the animal and clinical related field currently.A growing number of epidemiological studies have shown that MEHP associated with widespread disease,but the mechanism of cell survival under the stress of MEHP is not clear.In recent years,Saccharomyces cerevisiae gene deletion library is widely used in screening and identification of targets of different drugs in the cells and to explore the functioning pathways of drugs.The research of MEHP adaptability on S.cerevisiae genome level will contribute to the study of mammals gene and the related signaling pathways.In order to study the molecular mechanism of eukaryotic cells in terms of adapting MEHP stress,this topic have screened the related genes under MEHP stress by using the gene deletion mutant library of the eukaryotic model organism,S.cerevisiae.A total of 96 genes that involved in the MEHP-sensitivity were identified.These genes are involved in many cellular function,including ergosterol biosynthesis pathway,vacuolar protein sorting pathway and functions related to vacuolar ATPase.The high performance liquid chromatography is used to measure the intracellular MEHP concentrations of these 96 gene deletion mutants in response to MEHP stress.As compared to the wild type,49 mutants showed higher,and 21 mutants showed lower,intracellular concentrations of MEHP.The MEHP-sensitive phenotypes of erg2 and erg4 mutants can be partially suppressed by supplemented ergosterol or cholesterol in the medium.Ergosterol contents of of erg deletion mutants were not detected.These suggest that the function of ergosterol in S.cerevisiae are important for MEHP tolerance.In addition,the level of gene transcription related to the calcium/calcineurin signaling pathway was measured.Results indicate that MEHP can significantly inhibited the PMC1 transcription,this suggests that the calcium/calcineurin signaling pathway may be associated with the MEHP stress response.The results provide a basis for further understanding of MEHP functioning targets and mechanism in eukaryotic cells.