Rossi L, Hohn B, Tinland B
Friedrich Miescher-Institut, Basel, Switzerland.
Mol Gen Genet. 1993 Jun;239(3):345-53. doi: 10.1007/BF00276932.
Agrobacterium tumefaciens is able to transfer a piece of DNA, the T-DNA, to the nucleus of the plant cell. The VirD2 protein is required for the production of the T-DNA, it is tightly linked to the T-DNA and it is thought to direct it to the plant genome. Two nuclear localization signals (NLS), one in the N-terminal part and one in the C-terminal part of the VirD2 protein, have been shown to be able to target marker proteins to the plant nucleus. Here we analyze nuclear entry of the T-DNA complex using a new and very sensitive assay for T-DNA transfer. We show that optimal T-DNA transfer requires the VirD2 NLS located in the C-terminal part of the protein, whereas mutations in the N-terminal NLS coding sequence seem to have no effect on T-DNA transfer.
根癌土壤杆菌能够将一段DNA(即T-DNA)转移至植物细胞的细胞核。VirD2蛋白是产生T-DNA所必需的,它与T-DNA紧密相连,并被认为能将其导向植物基因组。已证明VirD2蛋白的两个核定位信号(NLS),一个位于N端部分,另一个位于C端部分,能够将标记蛋白靶向植物细胞核。在此,我们使用一种新的且非常灵敏的T-DNA转移检测方法,分析T-DNA复合体的核进入情况。我们发现,最佳的T-DNA转移需要位于蛋白C端部分的VirD2 NLS,而N端NLS编码序列中的突变似乎对T-DNA转移没有影响。