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Characterization of Multi-Functional Properties and ConformationalAnalysis of MutS2 from Thermotoga maritima MSB8
- Source :
- PLOS ONE(7): 4, PLoS ONE, Vol 7, Iss 4, p e34529 (2012), PLoS ONE
- Publication Year :
- 2012
-
Abstract
- The MutS2 homologues have received attention because of their unusual activities that differ from those of MutS. In this work, we report on the functional characteristics and conformational diversities of Thermotoga maritima MutS2 (TmMutS2). Various biochemical features of the protein were demonstrated via diverse techniques such as scanning probe microscopy (SPM), ATPase assays, analytical ultracentrifugation, DNA binding assays, size chromatography, and limited proteolytic analysis. Dimeric TmMutS2 showed the temperature-dependent ATPase activity. The non-specific nicking endonuclease activities of TmMutS2 were inactivated in the presence of nonhydrolytic ATP (ADPnP) and enhanced by the addition of TmMutL. In addition, TmMutS2 suppressed the TmRecA-mediated DNA strand exchange reaction in a TmMutL-dependent manner. We also demonstrated that small-angle X-ray scattering (SAXS) analysis of dimeric TmMutS2 exhibited nucleotide- and DNA-dependent conformational transitions. Particularly, TmMutS2-ADPnP showed the most compressed form rather than apo-TmMutS2 and the TmMutS2-ADP complex, in accordance with the results of biochemical assays. In the case of the DNA-binding complexes, the stretched conformation appeared in the TmMutS2-four-way junction (FWJ)-DNA complex. Convergences of biochemical- and SAXS analysis provided abundant information for TmMutS2 and clarified ambiguous experimental results.
- Subjects :
- DNA, Bacterial
Models, Molecular
Protein Structure
DNA Repair
Hydrolases
Protein Conformation
DNA recombination
ATPase
Marine and Aquatic Sciences
lcsh:Medicine
Biochemistry
law.invention
Nuclease
Endonuclease
chemistry.chemical_compound
Protein structure
Bacterial Proteins
X-Ray Diffraction
law
Scattering, Small Angle
Thermotoga maritima
Nucleotide
Homologous Recombination
Protein Interactions
lcsh:Science
Biology
Adenosine Triphosphatases
chemistry.chemical_classification
Multidisciplinary
biology
Enzyme Classes
Small-angle X-ray scattering
lcsh:R
Proteins
DNA
biology.organism_classification
Enzymes
Nucleic acids
chemistry
Earth Sciences
biology.protein
Recombinant DNA
lcsh:Q
Research Article
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- PLOS ONE(7): 4, PLoS ONE, Vol 7, Iss 4, p e34529 (2012), PLoS ONE
- Accession number :
- edsair.doi.dedup.....67f76b989a622983300ebe76b24ba351