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Nucleic Acids Research Advance Access originally published online on October 11, 2007
Nucleic Acids Research 2007 35(20):6917-6923; doi:10.1093/nar/gkm823
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Nucleic Acids Research, 2007, Vol. 35, No. 20 6917-6923
© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Nucleic Acid Enzymes

Extent to which hairpin opening by the Artemis:DNA-PKcs complex can contribute to junctional diversity in V(D)J recombination

Haihui Lu1, Klaus Schwarz2 and Michael R. Lieber1,*

1Department of Pathology, Department of Biochemistry & Molecular Biology, Department of Biological Sciences and Department of Molecular Microbiology & Immunology, Norris Comprehensive Cancer Center, Los Angeles, CA, USA and 2Institute for Clinical Transfusion Medicine and Immunogenetics, Ulm and Institute for Transfusion Medicine, University Hospital Ulm, Ulm, Germany

* To whom correspondence should be addressed. Tel: +1 323 865 0568; Fax: +1 323 865 3019; Email: lieber{at}usc.edu Correspondence may also be addressed to Klaus Schwarz. Tel: +49 731 150 642; Fax: +49 731 150 575; Email: klaus.schwarz{at}uni-ulm.de

Received July 31, 2007. Revised August 30, 2007. Accepted September 19, 2007.

V(D)J recombination events are initiated by cleavage at gene segments by the RAG1:RAG2 complex, which results in hairpin formation at the coding ends. The hairpins are opened by the Artemis:DNA-PKcs complex, and then joined via the nonhomologous DNA end joining (NHEJ) process. Here we examine the opening of the hairpinned coding ends from all of the 39 functional human VH elements. We find that there is some sequence-dependent variation in the efficiency and even the position of hairpin opening by Artemis:DNA-PKcs. The hairpin opening efficiency varies over a 7-fold range. The hairpin opening position varies over the region from 1 to 4 nt 3' of the hairpin tip, leading to a 2–8 nt single-stranded 3' overhang at each coding end. This information provides greater clarity on the extent to which the hairpin opening position contributes to junctional diversification in V(D)J recombination.


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