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

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Abstract

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.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Faculty of Biology, National and Kapodistrian University of Athens, Panepistimiopolis-Zographou, 15784, Athens, Greece

    Fotini Orfaniotou, Eleni Stefanidi, Athanassios Zees, Effrosini Boutou & Constantinos E. Vorgias

  2. Biomolecular Physics Laboratory, I/R-RP, National Centre for Scientific Research “Demokritos”, Aghia Paraskevi, 15310, Athens, Greece

    Pavlos Tzamalis, Angelos Thanassoulas & George Nounesis

  3. Institute of Biology, National Centre for Scientific Research “Demokritos”, Aghia Paraskevi, 15310, Athens, Greece

    Metaxia Vlassi

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  1. Fotini Orfaniotou
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  2. Pavlos Tzamalis
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  3. Angelos Thanassoulas
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  4. Eleni Stefanidi
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  7. Metaxia Vlassi
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  9. Constantinos E. Vorgias
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Correspondence to Constantinos E. Vorgias.

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F. Orfaniotou and P. Tzamalis contributed equally to this work.

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Orfaniotou, F., Tzamalis, P., Thanassoulas, A. et al. The stability of the archaeal HU histone-like DNA-binding protein from Thermoplasma volcanium . Extremophiles 13, 1–10 (2009). https://doi.org/10.1007/s00792-008-0190-6

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  • DOI: https://doi.org/10.1007/s00792-008-0190-6

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