Dumas F, Duckely M, Pelczar P, Van Gelder P, Hohn B
Department of Structural Biology, Biozentrum, Klingelbergstrasse 50, CH-4056 Basel, Switzerland; and Friedrich Miescher Institut, P.O. Box 2543, CH-4002 Basel, Switzerland.
Proc Natl Acad Sci U S A. 2001 Jan 16;98(2):485-90. doi: 10.1073/pnas.98.2.485. Epub 2001 Jan 9.
Transferred DNA (T-DNA) transfer from Agrobacterium tumefaciens into eukaryotic cells is the only known example of interkingdom DNA transfer. T-DNA is a single-stranded segment of Agrobacterium's tumor-inducing plasmid that enters the plant cell as a complex with the bacterial virulence proteins VirD2 and VirE2. The VirE2 protein is highly induced on contact of A. tumefaciens with a plant host and has been reported to act in late steps of transfer. One of its previously demonstrated functions is binding to the single-stranded (ss) T-DNA and protecting it from degradation. Recent experiments suggest other functions of the protein. A combination of planar lipid bilayer experiments, vesicle swelling assays, and DNA transport experiments demonstrated that VirE2 can insert itself into artificial membranes and form channels. These channels are voltage gated, anion selective, and single-stranded DNA-specific and can facilitate the efficient transport of single-stranded DNA through membranes. These experiments demonstrate a VirE2 function as a transmembrane DNA transporter, which could have applications in gene delivery systems.
从根癌农杆菌转移至真核细胞的转移DNA(T-DNA)是已知唯一的跨王国DNA转移实例。T-DNA是农杆菌致瘤质粒的单链片段,它作为与细菌毒力蛋白VirD2和VirE2的复合物进入植物细胞。VirE2蛋白在根癌农杆菌与植物宿主接触时高度诱导表达,据报道其在转移后期发挥作用。其先前已证实的功能之一是与单链(ss)T-DNA结合并保护其不被降解。最近的实验表明该蛋白还有其他功能。平面脂质双层实验、囊泡膨胀测定和DNA转运实验相结合表明,VirE2可插入人工膜并形成通道。这些通道是电压门控的、阴离子选择性的且对单链DNA具有特异性,能够促进单链DNA通过膜的高效转运。这些实验证明了VirE2作为跨膜DNA转运蛋白的功能,这在基因递送系统中可能具有应用价值。