亚砷酸钠染毒对酿酒酵母BY4742细胞骨架的影响

Effects of Sodium Arsenic on Cytoskeleton of Saccharomyces Cerevisiae BY4742

目的:利用酿酒酵母BY4742作为模式生物,探索不同浓度亚砷酸钠染毒后,细胞中砷的富集情况及对细胞生长和细胞骨架的影响。方法:分别以0.00 mmol/L、0.10 mmol/L、0.25 mmol/L、0.50 mmol/L、1.00 mmol/L和2.00 mmol/L亚砷酸钠处理酵母BY4742细胞,通过液体培养法和斑点法检测酵母细胞生存情况,利用双通道原子荧光光度计检测细胞内砷含量。此外,利用鬼笔环肽对细胞骨架进行染色并观察,qPCR检测细胞骨架相关基因表达变化量。结果:在0.10 mmol/L、0.25 mmol/L、0.50 mmol/L、1.00 mmol/L、2.00 mmol/L亚砷酸钠胁迫下,BY4742菌株生长受到不同程度的抑制,相对生长率显著降低为对照组的71%、63%、39%和6%。酵母细胞内砷含量随亚砷酸钠处理浓度的增加而增加,随着培养时间的延长细胞内砷排出胞外。此外,与对照组相比,以1.00 mmol/L和2.00 mmol/L亚砷酸钠进行染毒,细胞骨架出现斑块异常结构。以1.00 mmol/L亚砷酸钠进行染毒,arp2、arc15、arc19、arc18、las17、pfy1、cof1和crn1基因转录水平分别下调为对照组的59%、22%、28%、45%、28%、29%、35%和52%。结论:亚砷酸钠染毒后,砷在细胞内富集,抑制酵母BY4742菌株生长,抑制细胞骨架相关基因表达,导致其功能异常。

Objective:Using Saccharomyces cerevisiae BY4742 as model organism,to explore the arsenic enrichment conditions in cells and its influence on cell growth and cell skeleton after the different concentrations of sodium arsenite infected.Methods:BY4742 is treated with 0.00 mmol/L,0.10 mmol/L,0.25 mmol/L,0.50 mmol/L,1.00 mmol/L,2.00 mmol/L sodium arsenite,The survival of yeast cells is detected by liquid culture method and spot method,and the arsenic content in cells is detected by dual channel atomic fluorescence spectrometer.In addition,the cytoskeleton is stained with phalloidin and observed,and the expression of cytoskeleton related genes is detected by qPCR.Results:Under the 0.10 mmol/L,0.25 mmol/L,0.50 mmol/L,1.00 mmol/L,2.00 mmol/L sodium arsenite treatment,the growth of BY4742 yeast strain is inhibited in varying degrees,and the relative growth rate significantly decrease to 71%,63%,39%,and 6%.Further,arsenic in yeast cells increase with the increase of concentration of sodium arsenite treatment concentration.The arsenic content in yeast cells is excreted out of cells with the extension of culture time.Compared with control group,treatment with 1.00 mmol/L,2.00 mmol/L sodium arsenite,abnormal plaques cytoskeleton structure occured.Treatment with 1.00 mmol/L sodium arsenite,the level of arp2、arc15、arc19、arc18、las17、pfy1、cof1 and crn1 gene transcription decreased to 59%,22%,28%,45%,28%,29%,35%and 52%.Conclusion:Arsenic accumulates in cells after sodium arsenite exposure,inhibiting the BY4742 cell growth,and reducing the expression of cytoskeleton related genes,leading to its abnormal function.