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International Immunology, Vol. 12, No. 12, 1705-1713, December 2000
© 2000 Japanese Society for Immunology

A DNase I hypersensitive site near the murine {gamma}1 switch region contributes to insertion site independence of transgenes and modulates the amount of transcripts induced by CD40 ligation

Kristine Adams, Heidi Ackerly, Kirk Cunningham and Wesley Dunnick

Department of Microbiology and Immunology, University of Michigan Medical School, Room 6746, Medical Science Building II, 1301 East Catherine, Ann Arbor, MI 48109-0620, USA

Correspondence to: W. Dunnick

Several cis-acting elements regulate the expression of germline transcripts of heavy chain constant region genes and their subsequent switch recombination. To study such elements in the murine {gamma}1 gene, we have utilized a transgenic approach. In this study we focused on a DNase I hypersensitive site (termed `Site II') that lies about 2 kb 3' of the {gamma}1 promoter region and I exon, just 5' to the {gamma}1 switch region. We have reported that {gamma}1 transgenes with Site II display the characteristics of a locus control region (LCR) in that they are insertion site independent and copy number dependent. For the present study we prepared six lines of transgenic mice that have the promoter region and I exon, but lack Site II. Expression of RNA from {gamma}1 transgenes that lack Site II is not correlated with transgene copy number; expression is insertion site dependent. This result indicates that DNase hypersensitive Site II is an important part of the LCR-like elements in the murine {gamma}1 gene. RNA expression from the {gamma}1 transgenes that lack Site II is inducible by IL-4 and by CD40 ligation. However, the induction of transgenic RNA expression by CD40 ligation is greater than expected, suggesting that elements within Site II participate in negative regulation of the amount of germline transcripts after CD40 ligation.

Keywords: CD40 ligand, gene rearrangement, IL-4, isotype switching

The first two authors contributed equally to this work

Transmitting editor: K. Knight


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