Yoon J H, Love D C, Guhathakurta A, Hanover J A, Dhar R
Basic Research Laboratory, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
Mol Cell Biol. 2000 Dec;20(23):8767-82. doi: 10.1128/MCB.20.23.8767-8782.2000.
We identified the Schizosaccharomyces pombe mex67 gene (spmex67) as a multicopy suppressor of rae1-167 nup184-1 synthetic lethality and the rae1-167 ts mutation. spMex67p, a 596-amino-acid-long protein, has considerable sequence similarity to the Saccharomyces cerevisiae Mex67p (scMex67p) and human Tap. In contrast to scMEX67, spmex67 is essential for neither growth nor nuclear export of mRNA. However, an spmex67 null mutation (Deltamex67) is synthetically lethal with the rae1-167 mutation and accumulates poly(A)(+) RNA in the nucleus. We identified a central region (149 to 505 amino acids) within spMex67p that associates with a complex containing Rae1p that complements growth and mRNA export defects of the rae1-167 Deltamex67 synthetic lethality. This region is devoid of RNA-binding, N-terminal nuclear localization, and the C-terminal nuclear pore complex-targeting regions. The (149-505)-green fluorescent protein (GFP) fusion is found diffused throughout the cell. Overexpression of spMex67p inhibits growth and mRNA export and results in the redistribution of the diffused localization of the (149-505)-GFP fusion to the nucleus and the nuclear periphery. These results suggest that spMex67p competes for essential mRNA export factor(s). Finally, we propose that the 149-505 region of spMex67p could act as an accessory factor in Rae1p-dependent transport and that spMex67p participates at various common steps with Rae1p export complexes in promoting the export of mRNA.
我们鉴定出粟酒裂殖酵母mex67基因(spmex67)是rae1-167 nup184-1合成致死性以及rae1-167温度敏感突变的多拷贝抑制子。spMex67p是一种由596个氨基酸组成的蛋白质,与酿酒酵母Mex67p(scMex67p)和人类Tap具有相当的序列相似性。与scMEX67不同,spmex67对于生长和mRNA的核输出均非必需。然而,spmex67缺失突变(Deltamex67)与rae1-167突变是合成致死的,并在细胞核中积累多聚腺苷酸(+)RNA。我们在spMex67p中鉴定出一个中央区域(第149至505个氨基酸),该区域与一个包含Rae1p的复合物相关联,该复合物可弥补rae1-167 Deltamex67合成致死性的生长和mRNA输出缺陷。该区域缺乏RNA结合、N端核定位和C端核孔复合体靶向区域。(149-505)-绿色荧光蛋白(GFP)融合蛋白在整个细胞中呈弥散分布。spMex67p的过表达会抑制生长和mRNA输出,并导致(149-505)-GFP融合蛋白的弥散定位重新分布到细胞核和核周边。这些结果表明spMex67p竞争必需的mRNA输出因子。最后,我们提出spMex67p的149-505区域可能作为Rae1p依赖性转运中的辅助因子,并且spMex67p在促进mRNA输出的过程中与Rae1p输出复合体在多个共同步骤中发挥作用。