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Published July 4, 2000 | Published
Journal Article Open

V(D)J recombination is not activated by demethylation of the kappa locus

Abstract

V(D)J recombination is thought to be regulated by changes in the accessibility of target sites, such as modulation of methylation. To test whether demethylation of the kappa locus can activate recombination, we generated two recombinationally active B cell lines in which the gene for maintenance of genomic DNA methylation, Dnmt1, was flanked with loxP sites. Transduction with a retrovirus expressing both the cre recombinase and green fluorescent protein allowed us to purify recombinationally active cells devoid of methylation. Loss of methylation of the kappa locus was not sufficient to activate recombination, although transcription was activated in one line. It appears that demethylation of the kappa locus is not the rate-limiting step for altering accessibility and thus regulated demethylation does not generate specificity of recombination.

Additional Information

© 2000 The National Academy of Sciences. Contributed by David Baltimore, May 15, 2000. We thank members of the Jaenisch and Baltimore labs for advice and discussions. We are grateful to Dr. Luk van Parijs for advice on retroviral infections; Dr. Christopher Roman for assistance in generating Abelson lines and for discussions and comments on the manuscript; and Glen Paradis for fluorescence-activated cell sorting and technical assistance. D.B. is supported by a National Institutes of Health grant.

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