Journal article
Characterization of the recombination activities of the Entamoeba histolytica Rad51 recombinase
Molecular and biochemical parasitology, v 210(01-Feb)
Nov 2016
PMCID: PMC5125864
PMID: 27678398
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
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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.
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Details
- Title
- Characterization of the recombination activities of the Entamoeba histolytica Rad51 recombinase
- Creators
- Andrew A. Kelso - Clemson UniversitySteven D. Goodson - Clemson UniversitySuchitra Chavan - Clemson UniversityAmanda F. Say - Clemson UniversityAudrey Turchick - Clemson UniversityDeepti Sharma - Clemson UniversityLeAnna L. Ledford - Clemson UniversityErin Ratterman - Clemson UniversityKristin Leskoske - Clemson UniversityAda V. King - Clemson UniversityChristopher C. Attaway - Clemson UniversityYura Bandera - Clemson UniversityStephen H. Foulger - Clemson UniversityAlexander V. Mazin - Drexel UniversityLesly A. Temesvari - Clemson UniversityMichael G. Sehorn - Clemson University
- Publication Details
- Molecular and biochemical parasitology, v 210(01-Feb)
- Publisher
- Elsevier
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Biochemistry and Molecular Biology
- Web of Science ID
- WOS:000390823400011
- Scopus ID
- 2-s2.0-84998679657
- Other Identifier
- 991019168295804721
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- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Biochemistry & Molecular Biology
- Parasitology