Brown J A, Bharathi A, Ghosh A, Whalen W, Fitzgerald E, Dhar R
Laboratory of Molecular Virology, NCI, National Institutes of Health, Bethesda, Maryland 20892, USA.
J Biol Chem. 1995 Mar 31;270(13):7411-9. doi: 10.1074/jbc.270.13.7411.
A collection of fission yeast Schizosaccharomyces pombe conditional mutants was screened for defective nucleocytoplasmic transport of poly(A)+ RNA by fluorescence in situ hybridization. We identified a temperature-sensitive mutant that accumulated poly(A)+ RNA in the nucleus and have named it rae1-1, for ribonucleic acid export. All rae1-1 cells exhibit the defect in poly(A)+ RNA export within 30 min following a shift to the non-permissive temperature. In addition, in the rae1-1 mutant, actin and tubulin become disorganized, and cells undergo an irreversible cycle arrest. Results from experiments in which rae1-1 cells were arrested in various phases of the cell division cycle and then shifted to nonpermissive temperature suggest that cells are particularly vulnerable to loss of rae1 function during G2/M. However, the inability to export RNA from the nucleus to the cytoplasm was not limited to a particular phase of the cell division cycle. The rae1 gene was isolated by complementation and encodes a predicted protein of 352 amino acids with four beta-transducin/WD40 repeats.
通过荧光原位杂交技术,对裂殖酵母粟酒裂殖酵母的一组条件突变体进行筛选,以寻找聚腺苷酸(poly(A)+)RNA核质运输缺陷的突变体。我们鉴定出一个温度敏感型突变体,该突变体在细胞核中积累了poly(A)+ RNA,并将其命名为rae1-1,意为核糖核酸输出(ribonucleic acid export)。所有rae1-1细胞在转移到非允许温度后30分钟内,均表现出poly(A)+ RNA输出缺陷。此外,在rae1-1突变体中肌动蛋白和微管蛋白变得紊乱,细胞经历不可逆的周期阻滞。将rae1-1细胞阻滞在细胞分裂周期的各个阶段,然后转移到非允许温度的实验结果表明,细胞在G2/M期对rae1功能丧失尤为敏感。然而,无法将RNA从细胞核输出到细胞质并不局限于细胞分裂周期的特定阶段。通过互补作用分离出rae1基因,其编码一个预测的由352个氨基酸组成的蛋白质,含有四个β-转导蛋白/WD40重复序列。