Interactions of the DNA polymerase X from African Swine Fever Virus with the ssDNA. Properties of the total DNA-binding site and the strong DNA-binding subsite

Maria J. Jezewska, Michal R. Szymanski, Wlodzimierz Bujalowski

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Interactions of the polymerase X from the African Swine Fever Virus with the ssDNA have been studied, using quantitative fluorescence titration and fluorescence resonance energy transfer techniques. The primary DNA-binding subsite of the enzyme, independent of the DNA conformation, is located on the C-terminal domain. Association of the bound DNA with the catalytic N-terminal domain finalizes the engagement of the total DNA-binding site of the enzyme and induces a large topological change in the structure of the bound ssDNA. The free energy of binding includes a conformational transition of the protein. Large positive enthalpy changes accompanying the ASFV pol X-ssDNA association indicate that conformational changes of the complex are induced by the engagement of the N-terminal domain. The enthalpy changes are offset by large entropy changes accompanying the DNA binding to the C-terminal domain and the total DNA-binding site, predominantly resulting from the release of water molecules.

Original languageEnglish (US)
Pages (from-to)26-37
Number of pages12
JournalBiophysical Chemistry
Volume158
Issue number1
DOIs
StatePublished - Sep 2011

Fingerprint

African Swine Fever Virus
Viruses
Binding Sites
DNA
Catalytic DNA
Nucleic Acid Conformation
Fluorescence Resonance Energy Transfer
Enthalpy
Entropy
Enzymes
Fluorescence
Titration
Free energy
Conformations
DNA polymerase X
Water
Association reactions
Proteins
Molecules

Keywords

  • DNA replication
  • Fluorescence titrations
  • Motor protein
  • Polymerase
  • Protein-DNA interaction

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Organic Chemistry

Cite this

Interactions of the DNA polymerase X from African Swine Fever Virus with the ssDNA. Properties of the total DNA-binding site and the strong DNA-binding subsite. / Jezewska, Maria J.; Szymanski, Michal R.; Bujalowski, Wlodzimierz.

In: Biophysical Chemistry, Vol. 158, No. 1, 09.2011, p. 26-37.

Research output: Contribution to journalArticle

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