Ward B M, Lazarowitz S G
Department of Microbiology, University of Illinois, Urbana, Illinois 61801, USA.
Plant Cell. 1999 Jul;11(7):1267-76. doi: 10.1105/tpc.11.7.1267.
The nuclear export of proteins and RNAs has been studied in heterokaryons or by microinjecting test substrates into nuclei of HeLa cells or Xenopus oocytes. We have previously shown that the two movement proteins BR1 and BL1 encoded by the plant pathogenic squash leaf curl virus act in a coordinated manner to facilitate virus cell-to-cell movement and that one of these (BR1) is a nuclear shuttle protein. By using a novel in vivo cell-based assay for nuclear export in which nuclear-localized BR1 is trapped by BL1 and redirected to the cortical cytoplasm, we demonstrate that residues 177 to 198 of BR1 contain a leucine-rich nuclear export signal (NES) of the type found in the Rev protein encoded by the human immunodeficiency virus and in Xenopus TFIIIA. We further show that the TFIIIA NES can functionally replace the NES of BR1 in both nuclear export and viral infectivity. These findings suggest that this basic pathway for nuclear export is highly conserved among plant and animal cells and in yeast.
蛋白质和RNA的核输出已在异核体中进行研究,或通过将测试底物显微注射到HeLa细胞或非洲爪蟾卵母细胞的细胞核中来进行研究。我们之前已经表明,植物致病性南瓜曲叶病毒编码的两种运动蛋白BR1和BL1协同作用,促进病毒在细胞间移动,并且其中一种(BR1)是核穿梭蛋白。通过使用一种基于体内细胞的新型核输出检测方法,在该方法中,核定位的BR1被BL1捕获并重新导向皮质细胞质,我们证明BR1的177至198位残基包含一种富含亮氨酸的核输出信号(NES),这种类型的信号存在于人类免疫缺陷病毒编码的Rev蛋白和非洲爪蟾TFIIIA中。我们进一步表明,TFIIIA NES在核输出和病毒感染性方面都可以在功能上替代BR1的NES。这些发现表明,这种核输出的基本途径在植物、动物细胞和酵母中高度保守。