, 13:1 | Cite as

The stability of the archaeal HU histone-like DNA-binding protein from Thermoplasma volcanium

  • Fotini Orfaniotou
  • Pavlos Tzamalis
  • Angelos Thanassoulas
  • Eleni Stefanidi
  • Athanassios Zees
  • Effrosini Boutou
  • Metaxia Vlassi
  • George Nounesis
  • Constantinos E. Vorgias
Original Paper


The complete genome analysis of the archaeon Thermoplasma volcanium has revealed a gene assigned to encode the histone-like DNA-binding protein HU. Thermoplasma volcanium is a moderate thermophile growing around 60°C and it is adaptable to aerobic and anaerobic environment and therefore it is unique as a candidate for the origin of eukaryotic nuclei in the endosymbiosis hypothesis. The HU protein is the major component of the bacterial nuclei and therefore it is an important protein to be studied. The gene for HUTvo protein (huptvo) was cloned from the genomic DNA of T. volcanium and overexpressed in Escherichia coli. A fast and efficient purification scheme was established to produce an adequate amount of bioactive protein for biochemical and biophysical studies. Highly purified HUTvo was studied for its DNA-binding activity and thermostability. As studied by circular dichroism and high-precision differential scanning microcalorimetry, the thermal unfolding of HUTvo protein is reversible and can be well described by a two-state model with dissociation of the native dimeric state into denatured monomers. The ∆G versus T profile for HUTvo compared to the hyperthermophilic marine eubacterial counterpart from Thermotoga maritima, HUTmar, clearly shows that the archaeal protein has adopted a less efficient molecular mechanism to cope with high temperature. The molecular basis of this phenomenon is discussed.


Archaea Thermoplasma volcanium DNA-binding protein HU Cloning Purification Thermodynamic stability Thermal unfolding 


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Copyright information

© Springer 2008

Authors and Affiliations

  • Fotini Orfaniotou
    • 1
  • Pavlos Tzamalis
    • 2
  • Angelos Thanassoulas
    • 2
  • Eleni Stefanidi
    • 1
  • Athanassios Zees
    • 1
  • Effrosini Boutou
    • 1
  • Metaxia Vlassi
    • 3
  • George Nounesis
    • 2
  • Constantinos E. Vorgias
    • 1
  1. 1.Department of Biochemistry and Molecular Biology, Faculty of BiologyNational and Kapodistrian University of AthensAthensGreece
  2. 2.Biomolecular Physics Laboratory, I/R-RPNational Centre for Scientific Research “Demokritos”AthensGreece
  3. 3.Institute of BiologyNational Centre for Scientific Research “Demokritos”AthensGreece

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