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The stress-activated MAP kinase Sty1/Spc1 and a 3 '-regulatory element mediate UV-induced expression of the uvi15+ gene at the post-transcriptional level

机译:应力激活的MAP激酶Sty1 / Spc1和3'调控元件在转录后水平介导UV诱导的uvi15 +基因表达。

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摘要

Exposure of Schizosaccharomyces pombe cells to UV light results in increased uvi15+ gene expression at both the mRNA and protein levels, leading to elevated cell survival. This UV-induced expression of the uvi15+ gene was reduced in Δsty1 and Δwis1 cells lacking the stress-activated protein kinase pathway, but not in DNA damage checkpoint mutants. To further understand the cellular mechanisms responsible for this UV-induced expression, the transcription rate and mRNA half-life were investigated. Transcription run-on assays revealed that the rate of uvi15+ transcription was increased 1.8-fold regardless of Sty1 when cells were UV irradiated. The half-life of uvi15+ mRNA was also increased 1.5-fold after UV irradiation, but it was decreased in the Δsty1 background for both basal and UV-induced mRNAs, indicating that the stress-activated MAPK cascade can mediate UV-induced gene expression by increasing mRNA half-life. Deletion analyses identified a 54 nt element downstream of the distal poly(A) site, which was involved in the increased half-life of uvi15+ mRNA. These results suggest that both Sty1 and the 3'-regulatory element regulate UV-induced expression of the uvi15+ gene at the post-transcriptional level.
机译:粟酒裂殖酵母细胞暴露于紫外线下会导致mRNA和蛋白质水平的uvi15 +基因表达增加,从而导致细胞存活率提高。紫外线诱导的uvi15 +基因表达在缺乏应力激活蛋白激酶途径的Δsty1和Δwis1细胞中降低,但在DNA损伤检查点突变体中却没有降低。为了进一步了解负责此UV诱导表达的细胞机制,研究了转录速率和mRNA半衰期。转录连续测定显示,当细胞受到紫外线照射时,无论Sty1为何,uvi15 +转录的速率都会增加1.8倍。紫外线照射后,uvi15 + mRNA的半衰期也增加了1.5倍,但在基础和紫外线诱导的mRNA的Δsty1背景下均降低了,表明应激激活的MAPK级联可以介导紫外线诱导的基因表达。增加mRNA的半衰期。缺失分析鉴定了远端poly(A)位点下游的一个54 nt元件,该元件与uvi15 + mRNA的半衰期增加有关。这些结果表明,Sty1和3'调节元件均在转录后水平上调节UV诱导的uvi15 +基因表达。

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