The Journal of Experimental Medicine
ThymUS '08
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Published online November 19, 2007
doi:10.1084/jem.20071289
The Journal of Experimental Medicine, Vol. 204, No. 12, 3017-3026
The Rockefeller University Press, 0022-1007 $30.00
© 2007 Guikema et al.
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ARTICLE

APE1- and APE2-dependent DNA breaks in immunoglobulin class switch recombination

Jeroen E.J. Guikema1, Erin K. Linehan1, Daisuke Tsuchimoto2, Yusaku Nakabeppu2, Phyllis R. Strauss3, Janet Stavnezer1, and Carol E. Schrader1

1 Department of Molecular Genetics and Microbiology, Program in Immunology and Virology, University of Massachusetts Medical School, Worcester, MA 01655
2 Department of Immunobiology and Neuroscience, Medical Institute of Bioregulation, Kyushu University, Higashi-ku, Fukuoka 812-8582, Japan
3 Department of Biology, Northeastern University, Boston, MA 02115

CORRESPONDENCE Janet Stavnezer: Janet.Stavnezer{at}umassmed.edu

Antibody class switch recombination (CSR) occurs by an intrachromosomal deletion requiring generation of double-stranded breaks (DSBs) in switch-region DNA. The initial steps in DSB formation have been elucidated, involving cytosine deamination by activation-induced cytidine deaminase and generation of abasic sites by uracil DNA glycosylase. However, it is not known how abasic sites are converted into single-stranded breaks and, subsequently, DSBs. Apurinic/apyrimidinic endonuclease (APE) efficiently nicks DNA at abasic sites, but it is unknown whether APE participates in CSR. We address the roles of the two major mammalian APEs, APE1 and APE2, in CSR. APE1 deficiency causes embryonic lethality in mice; we therefore examined CSR and DSBs in mice deficient in APE2 and haploinsufficient for APE1. We show that both APE1 and APE2 function in CSR, resulting in the DSBs necessary for CSR and thereby describing a novel in vivo function for APE2.


Abbreviations used: AID, activation-induced cytidine deaminase; APE, apurinic/apyrimidinic endonuclease; ARP, aldehyde-reactive probe; CSR, class switch recombination; DBL, APE double deficient; DS, double stranded; DSB, DS break; HRP, horseradish peroxidase; LM-PCR, ligation-mediated PCR; NER, nt excision repair; S, switch; SS, single stranded; SSB, SS break; UNG, uracil DNA glycosylase.


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