Characterization of the recombination activities of the Entamoeba histolytica Rad51 recombinase
Autor: | Audrey Turchick, Yura Bandera, Steven D. Goodson, Michael G. Sehorn, Erin Ratterman, Lesly A. Temesvari, Alexander V. Mazin, Andrew A. Kelso, Stephen H. Foulger, Kristin L. Leskoske, LeAnna L. Ledford, Christopher C. Attaway, Suchitra Chavan, Amanda F. Say, Ada V. King, Deepti Sharma |
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Rok vydání: | 2016 |
Předmět: |
0301 basic medicine
DNA Repair DNA repair Protozoan Proteins RAD51 4 4'-Diisothiocyanostilbene-2 2'-Disulfonic Acid Biology Article Entamoeba invadens Substrate Specificity law.invention 03 medical and health sciences chemistry.chemical_compound Entamoeba histolytica Adenosine Triphosphate Plasmid law parasitic diseases Homologous Recombination Molecular Biology Hydrolysis DNA biology.organism_classification Recombinant Proteins Enzyme Activation 030104 developmental biology chemistry Biochemistry Recombinant DNA Nucleic Acid Conformation Calcium Parasitology Rad51 Recombinase Carrier Proteins Homologous recombination Plasmids Protein Binding |
Zdroj: | Molecular and Biochemical Parasitology. 210:71-84 |
ISSN: | 0166-6851 |
Popis: | The protozoan parasite responsible for human amoebiasis is Entamoeba histolytica. An important facet of the life cycle of E. histolytica involves the conversion of the mature trophozoite to a cyst. This transition is thought to involve homologous recombination (HR), which is dependent upon the Rad51 recombinase. Here, a biochemical characterization of highly purified ehRad51 protein is presented. The ehRad51 protein preferentially binds ssDNA, forms a presynaptic filament and possesses ATP hydrolysis activity that is stimulated by the presence of DNA. Evidence is provided that ehRad51 catalyzes robust DNA strand exchange over at least 5.4 kilobase pairs. Although the homologous DNA pairing activity of ehRad51 is weak, it is strongly enhanced by the presence of two HR accessory cofactors, calcium and Hop2-Mnd1. The biochemical system described herein was used to demonstrate the potential for targeting ehRad51 with two small molecule inhibitors of human RAD51. We show that 4,4′-diisothiocyanostilbene-2,2′-disulfonic acid (DIDS) inhibited ehRad51 by interfering with DNA binding and attenuated encystation in Entamoeba invadens, while B02 had no effect on ehRad51 strand exchange activity. These results provide insight into the underlying mechanism of homology-directed DNA repair in E. histolytica. |
Databáze: | OpenAIRE |
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