The Protein Journal

, Volume 32, Issue 7, pp 526–532 | Cite as

Metal-Binding Ability of VIP1: A bZIP Protein in Arabidopsis thaliana

  • Daisuke Tsugama
  • Shenkui Liu
  • Tetsuo Takano


VirE2-interacting protein 1 (VIP1) is an Arabidopsis thaliana bZIP transcription factor which regulates pathogen responses and rehydration responses. VIP1 has transcriptional activation potential, DNA-binding ability, and a nuclear-cytoplasmic shuttling property. These functions are possibly regulated by cofactors and/or post-translational modifications. During an investigation of the functions of VIP1, we discovered that VIP1 can react with an Ni2+-activated derivative of horseradish peroxidase, HisProbe-HRP, suggesting that VIP1 can bind Ni2+. Using truncated versions and mutated versions of VIP1, the Ni2+-binding region was narrowed. Using VIP1 H145Q and H145R mutants, which have H → Q and H → R mutations at the amino acid position 145 of VIP1, a trihistidine site at the amino acid position 144–146 was confirmed to be responsible for the Ni2+-binding ability. Immobilized-metal affinity chromatography (IMAC) suggested that VIP1 can bind Zn2+ and Co2+ as well as Ni2+, which is consistent with the known metal-chelating property of polyhistidine. In IMAC, the levels of purified VIP1 were not significantly different between denaturing and non-denaturing conditions, suggesting that the trihistidine is located on the surface of the native form of VIP1. In gel shift assays, VIP1-dependent decreases of electrophoretic mobilities of DNA probes were further decreased by Co2+. Among wild-type VIP1 and the H145Q and H145R mutants, H145R was the least sensitive to the effect of Co2+ in the gel shift assays. These results suggest that the Co2+ and the metal-binding site of VIP1 affect the interaction between VIP1 and DNA.


Arabidopsis thaliana Transcription factor Polyhistidine DNA–protein interaction 



Nuclear localization signal


Nuclear export signal


Horseradish peroxidase


Tris-buffered saline


Immobilized-metal affinity chromatography



This work was supported by a Grant-in-aid for Scientific Research (22·2144) to D.T.


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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  1. 1.Asian Natural Environmental Science Center (ANESC)The University of TokyoNishitokyo-shiJapan
  2. 2.Alkali Soil Natural Environmental Science Center (ASNESC)Northeast Forestry UniversityHarbinPeople’s Republic of China

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