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DNA-PKcs regulates a single-stranded DNA endonuclease activity of Artemis |
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| Authors: | Jiafeng Gu Sicong Li Xiaoshan Zhang Ling-Chi Wang Doris Niewolik Klaus Schwarz Randy J Legerski Ebrahim Zandi Michael R Lieber |
| Institution: | 1. Department of Pathology, Norris Comprehensive Cancer Center, Los Angeles, CA, USA;2. Department of Biochemistry & Molecular Biology, Norris Comprehensive Cancer Center, Los Angeles, CA, USA;3. Department of Biological Sciences, Section on Molecular & Computational Biology, Los Angeles, CA, USA;4. Department of Molecular Microbiology & Immunology, Norris Comprehensive Cancer Center, Los Angeles, CA, USA;5. Department of Genetics, The University of Texas MD Anderson Cancer Center, University of Texas, 1515 Holcombe Boulevard, Houston, TX 77030, USA;6. Institute for Clinical Transfusion Medicine and Immunogenetics, Ulm, Germany;7. Institute for Transfusion Medicine, University of Ulm, Ulm, Germany;1. Quantum Beam Science Directorate, Japan Atomic Energy Agency, 8-1-7, Umemidai, Kizugawa-city 619-0215, Japan;2. Faculty of Life and Medical Sciences, Doshisha University,1-3 Tatara Miyakodani, Kyotanabe City 610-0394, Japan;1. Department of Molecular Biosciences, The University of Texas at Austin, Austin, TX 78712, USA;2. Howard Hughes Medical Institute, The University of Texas at Austin, Austin, TX 78712, USA;3. Center for Systems and Synthetic Biology, The University of Texas at Austin, Austin, TX 78712, USA |
| Abstract: | Human nuclease Artemis belongs to the metallo-beta-lactamase protein family. It acquires double-stranded DNA endonuclease activity in the presence of DNA-PKcs. This double-stranded DNA endonuclease activity is critical for opening DNA hairpins in V(D)J recombination and is thought to be important for processing overhangs during the nonhomologous DNA end joining (NHEJ) process. Here we show that purified human Artemis exhibits single-stranded DNA endonuclease activity. This activity is proportional to the amount of highly purified Artemis from a gel filtration column. The activity is stimulated by DNA-PKcs and modulated by purified antibodies raised against Artemis. Moreover, the divalent cation-dependence and sequence-dependence of this single-stranded endonuclease activity is the same as the double-stranded DNA endonuclease activity of Artemis:DNA-PKcs. These findings further expand the range of DNA substrates upon which Artemis and Artemis:DNA-PKcs can act. The findings are discussed in the context of NHEJ. |
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