Zolotukhin A S, Felber B K
Human Retrovirus Pathogenesis Group, ABL-Basic Research Program, National Cancer Institute-Frederick Cancer Research and Development Center, Frederick, Maryland 21702-1201, USA.
J Virol. 1999 Jan;73(1):120-7. doi: 10.1128/JVI.73.1.120-127.1999.
Human immunodeficiency virus type 1 (HIV-1) Rev contains a leucine-rich nuclear export signal that is essential for its nucleocytoplasmic export mediated by hCRM1. We examined the role of selected nucleoporins, which are located in peripheral structures of the nuclear pore complex and are thought to be involved in export, in Rev function in human cells. First, we found that upon actinomycin D treatment, Nup98, but not Nup214 or Nup153, is able to translocate to the cytoplasm of HeLa cells, demonstrating that Nup98 may act as a soluble factor. We further showed that Rev can recruit Nup98 and Nup214, but not Nup153, to the nucleolus. We also found that the isolated FG-containing repeat domains of Nup98 and Nup214, but not those of Nup153, competitively inhibit the Rev/RRE-mediated expression of HIV. Taken together, the recruitment of Nup98 and Nup214 by Rev and the competitive inhibition exhibited by their NP domains demonstrate direct participation of Nup98 and Nup214 in the Rev-hCRM1-mediated export.
1型人类免疫缺陷病毒（HIV-1）的Rev蛋白含有一个富含亮氨酸的核输出信号，该信号对于其由hCRM1介导的核质输出至关重要。我们研究了位于核孔复合体周边结构且被认为参与输出过程的特定核孔蛋白在人类细胞Rev功能中的作用。首先，我们发现用放线菌素D处理后，Nup98能够转移至HeLa细胞的细胞质中，而Nup214或Nup153则不能，这表明Nup98可能作为一种可溶性因子发挥作用。我们进一步表明，Rev能够将Nup98和Nup214招募至核仁，但不能招募Nup153。我们还发现，Nup98和Nup214分离出的含FG重复结构域能够竞争性抑制Rev/RRE介导的HIV表达，而Nup153的相应结构域则不能。综上所述，Rev对Nup98和Nup214的招募以及它们的NP结构域所表现出的竞争性抑制作用表明，Nup98和Nup214直接参与了Rev-hCRM1介导的输出过程。
