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Agrobacterium VirD2 protein interacts with plant host cyclophilins
1Department of Microbiology, Box 357242, University of Washington, Seattle, WA 98195-7242, USA.
Summary
Host cyclophilins interact with Agrobacterium tumefaciens VirD2 protein, crucial for T-DNA transfer during plant transformation. Cyclosporin A disrupts this interaction, inhibiting crown gall tumor formation.
Area of Science:
- Plant molecular biology
- Bacterial genetics
- Biochemistry
Background:
- Agrobacterium tumefaciens transfers T-complex DNA to plants, causing crown gall tumors.
- The T-complex includes T-DNA, VirD2 endonuclease, and VirE2 DNA-binding protein.
Purpose of the Study:
- Identify Arabidopsis thaliana proteins interacting with VirD2 and VirE2.
- Characterize the interaction between VirD2 and identified host proteins.
Main Methods:
- Yeast two-hybrid system screening for protein-protein interactions.
- In vitro biochemical assays to study VirD2-cyclophilin interaction disruption.
Main Results:
- Identified three VirD2- and two VirE2-interacting proteins in Arabidopsis.
- Characterized interaction of VirD2 with two Arabidopsis cyclophilin isoforms.
- Localized the VirD2 interaction domain away from catalytic and nuclear localization sites.
- Demonstrated cyclosporin A disrupts VirD2-cyclophilin interaction and inhibits Agrobacterium-mediated transformation.
Conclusions:
- Host cyclophilins are implicated in the T-DNA transfer process by Agrobacterium.
- VirD2-cyclophilin interaction is a potential target for modulating plant transformation efficiency.