Saavedra C, Tung K S, Amberg D C, Hopper A K, Cole C N
Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755, USA.
Genes Dev. 1996 Jul 1;10(13):1608-20. doi: 10.1101/gad.10.13.1608.
The response of eukaryotic cells to heat shock and other forms of stress occurs at both transcriptional and post-transcriptional levels. We used in situ hybridization to determine whether stress affected the subcellular distribution of poly(A)+ RNA in Saccharomyces cerevisiae. Following induction of stress by either heat shock (42 degrees C) or addition of a high concentration of ethanol (10%), the nucleocytoplasmic export of most poly(A)+ RNA was blocked. In situ hybridization indicated that heat-inducible SSA4 and SSA1 mRNAs were exported from nuclei under these same conditions. On the other hand, both GAL1 and URA3 transcripts expressed from the SSA4 promoter accumulated in nuclei following heat shock. Sequences within either the 5' 1600 or the 3' 500 nucleotides of SSA4 mRNA were sufficient to direct GAL1 mRNA to the cytoplasm during stress. The export of SSA4 mRNA following stress required functional nuclear pore complexes, as SSA4 mRNA accumulated in nuclei following heat shock of cells containing temperature-sensitive nucleoporins. However, the selective export of SSA4 mRNA was maintained in heat-shocked cells carrying temperature-sensitive alleles of RNA1, PRP20, or an inducible dominant-negative allele of GSP1, the S. cerevisiae homolog of RAN/TC4. The results reported here suggest that there is selective export of mRNA in yeast.
真核细胞对热休克和其他形式应激的反应发生在转录和转录后水平。我们使用原位杂交来确定应激是否会影响酿酒酵母中聚腺苷酸(poly(A)+)RNA的亚细胞分布。在通过热休克(42℃)或添加高浓度乙醇(10%)诱导应激后,大多数聚腺苷酸(poly(A)+)RNA的核质输出被阻断。原位杂交表明,在相同条件下,热诱导的SSA4和SSA1 mRNA从细胞核中输出。另一方面,在热休克后,从SSA4启动子表达的GAL1和URA3转录本都在细胞核中积累。SSA4 mRNA 5'端1600个核苷酸或3'端500个核苷酸内的序列足以在应激期间将GAL1 mRNA导向细胞质。应激后SSA4 mRNA的输出需要功能性核孔复合体,因为在含有温度敏感型核孔蛋白的细胞热休克后,SSA4 mRNA在细胞核中积累。然而,在携带RNA1、PRP20温度敏感等位基因或GSP1(酿酒酵母中RAN/TC4的同源物)的诱导型显性负等位基因的热休克细胞中,SSA4 mRNA的选择性输出得以维持。这里报道的结果表明酵母中存在mRNA的选择性输出。