Article
- The EMBO Journal (2004) 23, 1987 - 1997
- doi:10.1038/sj.emboj.7600206
Published online: 8 April 2004
Subject Categories:
Functional and biochemical dissection of the structure-specific nuclease ARTEMIS
Ulrich Pannicke1, Yunmei Ma2, Karl-Peter Hopfner3, Doris Niewolik1, Michael R Lieber2 and Klaus Schwarz1
- Institute for Clinical Transfusion Medicine and Immunogenetics, Ulm, Department of Transfusion Medicine, University of Ulm, Ulm, Germany
- Departments of Pathology, Biochemistry & Molecular Biology, Biological Sciences, and Molecular Microbiology & Immunology, Norris Comprehensive Cancer Center, Los Angeles, CA, USA
- Institute of Biochemistry and Gene Center, University of Munich, Munich, Germany
Correspondence to:
Klaus Schwarz, Institute for Clinical Transfusion Medicine, and Immunogenetics, Ulm, Department of Transfusion Medicine, University of Ulm, Helmholtzstrasse 10, 89081 Ulm, Germany. Tel.: +49 731 150 642; Fax: +49 731 150 575; E-mail: klaus.schwarz@medizin.uni-ulm.de
Michael R Lieber, Departments of Pathology, Biochemistry & Molecular Biology, Biological Sciences, and Molecular Microbiology & Immunology, Norris Comprehensive Cancer Center, Rm 5428, 1441 Eastlake Avenue, Los Angeles, CA 90033, USA. E-mail: lieber@usc.edu
Received 27 August 2003; Accepted 18 March 2004
Abstract
During V(D)J recombination, the RAG1 and RAG2 proteins form a complex and initiate the process of rearrangement by cleaving between the coding and signal segments and generating hairpins at the coding ends. Prior to ligation of the coding ends by DNA ligase IV/XRCC4, these hairpins are opened by the ARTEMIS/DNA-PKcs complex. ARTEMIS, a member of the metallo-
-lactamase superfamily, shares several features with other family members that act on nucleic acids. ARTEMIS exhibits exonuclease and, in concert with DNA-PKcs, endonuclease activities. To characterize amino acids essential for its catalytic activities, we mutated nine evolutionary conserved histidine and aspartic acid residues within ARTEMIS. Biochemical analyses and a novel in vivo V(D)J recombination assay allowed the identification of eight mutants that were defective in both overhang endonucleolytic and hairpin-opening activities; the 5' to 3' exonuclease activity of ARTEMIS, however, was not impaired by these mutations. These results indicate that the hairpin-opening activity of ARTEMIS and/or its overhang endonucleolytic activity are necessary but its exonuclease activity is not sufficient for the process of V(D)J recombination.
Keywords:
- ARTEMIS,
- metallo--lactamase,
- SCID,
- V(D)J recombination
MORE ARTICLES LIKE THIS
These links to content published by NPG are automatically generated
REVIEWS
The mechanisms of immune diversification and their disorders
Nature Reviews Immunology Review (01 Dec 2003)
Oncogene Review
NEWS AND VIEWS
Mre11: roles in DNA repair beyond homologous recombination
Nature Structural & Molecular Biology News and Views (01 Aug 2009)
Nature News and Views (18 Apr 2002)
RESEARCH
DNA-PK autophosphorylation facilitates Artemis endonuclease activity
The EMBO Journal Article (23 Aug 2006)
