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Mre11 and Rad50 from Pyrococcus furiosus: cloning and biochemical characterization reveal an evolutionarily conserved
K P Hopfner1, A Karcher, D Shin
1Department of Molecular Biology and Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
Journal of Bacteriology
|October 13, 2000
Summary
DNA double-strand break repair is vital. Researchers found heat-stable Mre11-Rad50 archaeal complex with nuclease activity, confirming its presence across all life kingdoms.
Area of Science:
- Molecular Biology
- Biochemistry
- Genetics
Background:
- DNA double-strand breaks are critical nucleic acid events.
- Deficiencies in DNA repair lead to severe phenotypes.
- The Mre11-Rad50 complex is central to eukaryotic double-strand break repair.
Purpose of the Study:
- To isolate and characterize Mre11 and Rad50 homologues from Pyrococcus furiosus.
- To investigate the biochemical properties of the archaeal Mre11-Rad50 complex.
Main Methods:
- Isolation of Mre11 and Rad50 proteins from Pyrococcus furiosus.
- Biochemical assays to determine nuclease activity and ATP dependence.
Main Results:
- A large, heat-stable complex of Mre11 and Rad50 was isolated.
- The complex exhibits single-strand endonuclease activity.
- ATP-dependent double-strand-specific exonuclease activity was demonstrated.
Conclusions:
- Confirms the identification and function of Pyrococcus furiosus Mre11 and Rad50 homologues.
- Demonstrates that functional Mre11-Rad50 complexes with similar biochemical properties exist in all life kingdoms.