Agrobacterium-delivered VirE2 interacts with host nucleoporin CG1 to facilitate the nuclear import of VirE2-coated T complex.
Agrobacterium-delivered VirE2 interacts with host nucleoporin CG1 to facilitate the nuclear import of VirE2-coated T complex.
复制标题
农杆菌递送的 VirE2 与宿主核孔蛋白 CG1 相互作用，促进 VirE2 包被的 T 复合物进入核。
DOI：
10.1073/pnas.200964517
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发表时间：
2020
影响因子：
11.1
通讯作者：
Pan, S.Q.
中科院分区：
文献类型：
--
作者：
Li, X.;Yang, Q.;Peng, L.;Tu, H.;Lee, L.-Y.;Gelvin, S.B.;Pan, S.Q.
根癌农杆菌引起根癌病之病原。该细菌能够在转化过程中将一段单链DNA（ssDNA）转移到受体细胞中，并且它已被广泛用作植物和非植物生物的遗传修饰工具。转移DNA（T-DNA）被认为是由两种毒力蛋白VirD 2和VirE 2作为靶向宿主细胞核的核蛋白复合物（T-complex）护送的。然而，目前尚不清楚这种拟议的大DNA-蛋白质复合物如何在自然环境中通过宿主核孔递送。在这里，我们研究了自然的核输入theAgrobacterium-delivered ssDNA结合蛋白VirE 2植物细胞内使用split-GFP的方法与新构建的T-DNA-free菌株。我们的研究结果表明，VirE 2靶向进入宿主细胞核中的VirD 2和T-DNA依赖的方式。与VirD 2结合到植物输入蛋白α进行核输入不同，VirE 2直接与宿主核孔复合体组分核孔蛋白CG 1相互作用，以促进其核摄取和转化过程。我们的数据表明，一个合作的核进口模型中，T-DNA被引导到主机核孔VirD 2和通过孔VirE 2和主机核孔蛋白CG 1之间的相互作用的援助。我们假设，这个大的线性核蛋白复合物（T-复合物）是有针对性的细胞核的“头”的VirD 2-importin相互作用的指导和进入细胞核的VirE 2-核孔蛋白相互作用的横向援助。
Agrobacterium tumefaciensis the causal agent of crown gall disease. The bacterium is capable of transferring a segment of single-stranded DNA (ssDNA) into recipient cells during the transformation process, and it has been widely used as a genetic modification tool for plants and nonplant organisms. Transferred DNA (T-DNA) has been proposed to be escorted by two virulence proteins, VirD2 and VirE2, as a nucleoprotein complex (T-complex) that targets the host nucleus. However, it is not clear how such a proposed large DNA–protein complex is delivered through the host nuclear pore in a natural setting. Here, we studied the natural nuclear import of theAgrobacterium-delivered ssDNA-binding protein VirE2 inside plant cells by using a split-GFP approach with a newly constructed T-DNA–free strain. Our results demonstrate that VirE2 is targeted into the host nucleus in a VirD2- and T-DNA–dependent manner. In contrast with VirD2 that binds to plant importin α for nuclear import, VirE2 directly interacts with the host nuclear pore complex component nucleoporin CG1 to facilitate its nuclear uptake and the transformation process. Our data suggest a cooperative nuclear import model in which T-DNA is guided to the host nuclear pore by VirD2 and passes through the pore with the assistance of interactions between VirE2 and host nucleoporin CG1. We hypothesize that this large linear nucleoprotein complex (T-complex) is targeted to the nucleus by a “head” guide from the VirD2–importin interaction and into the nucleus by a lateral assistance from the VirE2–nucleoporin interaction.