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PMID: 11149937 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

An Agrobacterium VirE2 channel for transferred-DNA transport into plant cells.

Dumas F, Duckely M, Pelczar P, Van Gelder P, Hohn B

Abstract

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.

MeSH Terms
Agrobacterium tumefaciens/physiology Anions/metabolism Arabidopsis/microbiology Bacterial Proteins/chemistry,physiology Biological Transport Cell Membrane Permeability DNA, Bacterial/genetics,metabolism DNA, Single-Stranded/genetics,metabolism DNA-Binding Proteins/chemistry,physiology Diffusion Ion Channel Gating/physiology Ion Channels Ion Transport Lipid Bilayers Macromolecular Substances Membrane Lipids/metabolism Membrane Proteins/chemistry,physiology Models, Biological Molecular Chaperones/metabolism Nuclear Proteins/metabolism Plant Proteins/metabolism Plasmids/genetics Protein Binding Recombinant Fusion Proteins/chemistry,metabolism Transformation, Bacterial/physiology Virulence Factors
Chemicals
Anions Bacterial Proteins DNA, Bacterial DNA, Single-Stranded DNA-Binding Proteins Ion Channels Lipid Bilayers Macromolecular Substances Membrane Lipids Membrane Proteins Molecular Chaperones Nuclear Proteins Plant Proteins Recombinant Fusion Proteins Virulence Factors
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Dumas F
Department of Structural Biology, Biozentrum, Klingelbergstrasse 50, CH-4056 Basel, Switzerland; and Friedrich Miescher Institut, P.O. Box 2543, CH-4002 Basel, Switzerland. fabricedumas@yahoo.fr
Duckely M
Pelczar P
Van Gelder P
Hohn B
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
2001-01-16
Epub
2001-00-09
Pages
485-90
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC14613
Subset
IM
Corrections
CommentIn
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