Elp3基因缺失对酿酒酵母咖啡因耐受性的影响

摘要：

过量咖啡因(Caffeine,CAF)对真核细胞有毒害作用.Elp3是组蛋白乙酰转移酶Elongator复合物的催化亚基,其缺失会影响Elongator的功能,进而使酵母产生对CAF敏感表型.为研究酿酒酵母CAF耐受性,以不同浓度CAF处理野生型、elp3⊿及转入完整和部分缺失Elp3的elp3⊿菌株,检测其敏感性,并通过RT-qPCR和ChIP实验研究了热激胁迫下SSA3的表达.结果发现常温下各菌株生长均随CAF浓度的增加而减缓,且elp3⊿细胞CAF耐受性下降更显著;高温处理使elp3⊿菌株对CAF更加敏感;在elp3⊿菌株中转入两保守功能域缺失的Elp3,其咖啡因耐受性并无明显变化;无论热激与否,elp3⊿菌株SSA3表达均显著低于野生型对照,Elp3直接参与热激胁迫下SSA3的转录延伸.研究表明Elp3基因缺失和高温处理均可使酿酒酵母的CAF耐受性降低;Elp3两保守功能域为其参与CAF胁迫反应所必需;高温协同处理下elp3⊿菌株CAF耐受性的显著降低可能与此条件下SSA3等应激基因表达受阻有关.

An excess of caffeine is cytotoxic to all eukaryotic cell types.Elp3 is the catalytic subunit of histone acetyltransferase Elongator complex,its absence or dysfunction may affect the function of the Elongator and result in the sensitive phenotype of yeast.To study the CAF tolerance of Saccharomyces cerevisiae,cell growth was detected in wild-type,elp3⊿and full length or partial deletion Elp3 transformed elp3⊿cells by sensitivity detection and growth curve drawing under different concentrations of CAF and/or high temperature stress conditions.SSA3 expression in wild-type and elp3⊿stains was detected by RT-qPCR and ChIP experiments under heat shock stress.The results of caffeine sensitivity detection and growth curve plotted show that the growth of all stains significantly slow down with the increase of CAF concentration,especially of elp3⊿cells.High temperature co-processing will increase their CAF sensitive phenotype.Both C and N terminal conserved domain deletion Elp3 have no compensation to the CAF sensitivity phenotype of elp3⊿strain.RT-PCRthan wild-type.ChIP experiments further confirmed that Elp3 directly involved in heat shock stress SSA3 gene transcription elongation.These suggest that both Elp3 deletion and high temperature treatment decrease the CAF tolerance of Saccharomyces cerevisiae,the two conserved domain of Elp3 are both essential to this function and the more sensitive phenotype displayed by elp3⊿strains under caffeine and high temperature stress comes from the reduction of stress genes such as SSA3 under stress conditions.

作者：

胡孟可 吴秀丽 李芬

Hu Mengke;Wu Xiuli;Li Fen(College of Life Sciences,Henan Normal University,Xinxiang 453007,China)

机构地区：

betway官方app 生命科学学院

出处：

《betway官方app 学报：自然科学版》 CAS 北大核心 2022年第2期143-149,共7页

Journal of Henan Normal University(Natural Science Edition)

基金：

国家自然科学基金(30600343) 河南省高校科技创新团队支持计划(13IRTSTHN009).

关键词：

咖啡因耐受性 酿酒酵母 ELP3 SSA3 保守结构域

caffeine tolerance Saccharomyces cerevisiae Elp3 SSA3 conserved domain

分类号：

Q291 [生物学—细胞生物学]

Elp3基因缺失对酿酒酵母咖啡因耐受性的影响.pdf