The disordered region of arabidopsis VIP1 binds the agrobacterium VirE2 protein outside its DNA-binding site

Michal Maes, Einav Amit, Tsafi Danieli, Mario Lebendiker, Abraham Loyter, Assaf Friedler*, Gideon Schreiber

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Agrobacterium is a pathogen that genetically transforms plants. The bacterial VirE2 protein envelopes the T-DNA of Agrobacterium and protects it from degradation. Within the transfected cells, VirE2 interacts with the plant VIP1 leading to nuclear transport of the T-DNA complex. Active VirE2 is an oligomer with a tendency to aggregate, hampering its studies at the molecular level. In addition, no structural or quantitative information is available regarding VIP1 or its interactions. The lack of information is mainly because both VIP1 and VirE2 are difficult to express and purify. Here, we present the development of efficient protocols that resulted in pure and stable Histagged VIP1 and VirE2. Circular dichroism spectroscopy and computational predictions indicated that VIP1 is mostly intrinsically disordered. This may explain the variety of protein-protein interactions it participates in. Size exclusion chromatography revealed that VirE2 exists in a two-state equilibrium between a monomer and an oligomeric form. Using the purified proteins, we performed peptide array screening and revealed the binding sites on both proteins. VirE2 binds the disordered regions of VIP1, while the site in VirE2 that binds VIP1 is different from the VirE2 DNA-binding site. Peptides derived from these sites may be used as lead compounds that block Agrobacterium infection of plants.

Original languageEnglish
Pages (from-to)439-446
Number of pages8
JournalProtein Engineering, Design and Selection
Volume27
Issue number11
DOIs
StatePublished - 1 Nov 2014

Bibliographical note

Publisher Copyright:
© The Author 2014. Published by Oxford University Press. All rights reserved.

Keywords

  • Agrobacterium
  • Peptide arrays
  • Protein expression and purification
  • VIP1
  • VirE2

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