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Nuclear export in plants. Use of geminivirus movement proteins for a cell-based export assay.

机译:植物核出口。双生病毒运动蛋白在基于细胞的出口检测中的用途。

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摘要

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.
机译:已经在异核体中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核中研究了蛋白质和RNA的核输出。先前我们已经表明,植物病原性南瓜叶片卷曲病毒编码的两个运动蛋白BR1和BL1以协调的方式起作用,以促进病毒细胞间的运动,其中一个(BR1)是核穿梭蛋白。通过使用一种新的基于体内细胞的核输出方法,其中核定位的BR1被BL1捕获并重定向到皮质细胞质,我们证明BR1的残基177至198包含富含亮氨酸的核输出信号(NES)在人类免疫缺陷病毒编码的Rev蛋白和非洲爪蟾TFIIIA中发现的类型。我们进一步表明,TFIIIA NES可以在核输出和病毒感染性方面取代BR1的NES。这些发现表明,这种核出口的基本途径在动植物细胞和酵母中高度保守。

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