Kim M, Lee W, Park J, Kim J B, Jang Y K, Seong R H, Choe S Y, Park S D
School of Biological Sciences, College of Natural Sciences, Seoul National University, Seoul 151-742, Republic of Korea.
Nucleic Acids Res. 2000 Sep 1;28(17):3392-402. doi: 10.1093/nar/28.17.3392.
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 Deltasty1 and Deltawis1 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 Deltasty1 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.
粟酒裂殖酵母细胞暴露于紫外线下会导致uv i15(+)基因在mRNA和蛋白质水平上的表达增加,从而提高细胞存活率。在缺乏应激激活蛋白激酶途径的Deltasty1和Deltawis1细胞中,uv i15(+)基因的这种紫外线诱导表达降低,但在DNA损伤检查点突变体中则没有降低。为了进一步了解负责这种紫外线诱导表达的细胞机制,研究了转录速率和mRNA半衰期。转录延伸分析表明,当细胞受到紫外线照射时,无论是否存在Sty1,uv i15(+)的转录速率都会增加1.8倍。紫外线照射后,uv i15(+) mRNA的半衰期也增加了1.5倍,但在Deltasty1背景下,基础mRNA和紫外线诱导的mRNA的半衰期均降低,这表明应激激活的MAPK级联反应可以通过增加mRNA半衰期来介导紫外线诱导的基因表达。缺失分析确定了远端多聚腺苷酸化位点下游的一个54 nt元件,该元件参与了uv i15(+) mRNA半衰期的增加。这些结果表明,Sty1和3'调控元件都在转录后水平上调节uv i15(+)基因的紫外线诱导表达。