The Journal of Experimental Medicine
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© The Rockefeller University Press, 0022-1007/1997/7/91/ $5.00
The Journal of Experimental Medicine, Volume 186, Number 1, July 7, 1997 91-100

Articles

Identification of a DNA Segment Exhibiting Rearrangement Modifying Effects upon Transgenic {delta}-deleting Elements

Karen M. Janowski*, Stephanie Ledbetter*, Matthew S. Mayo{ddagger}, and Richard D. Hockett, Jr.*

From the * Department of Pathology, and the {ddagger} Biostatistics Unit, Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama 35233-7331

Control of the rearrangement and expression of the T cell receptor {alpha} and {delta} chains is critical for determining T cell type. The process of {delta} deletion is a candidate mechanism for maintaining separation of the {alpha} and {delta} loci. Mice harboring a transgenic reporter {delta} deletion construct show {alpha}/β T cell lineage–specific use of the transgenic elements. A 48-basepair segment of DNA, termed HPS1A, when deleted from this reporter construct, loses tight lineage-specific rearrangement control of transgenic elements, with abundant rearrangements of transgenic {delta}-deleting elements now in {gamma}/{delta} T cells. Furthermore, HPS1A augments recombination frequency of extrachromosomal substrates in an in vitro recombination assay. DNA binding proteins recognizing HPS1A have been identified and are restricted to early B and T cells, during the time of active rearrangement of endogenous TCR and immunoglobulin loci. These data are consistent with {delta} deletion playing an important role in maintaining separate TCR {alpha} and {delta} loci.

Address correspondence to R.D. Hockett, Jr., UAB Dept. of Pathology, WP230, 618 S. 18th St., Birmingham, AL 35233. Phone: 205-934-6246; FAX: 205-975-7074; E-mail: Hockett{at}wp.path.uab.edu

1Abbreviations used in this paper: AMP, ampicillin; CAM, chloramphenicol; DTT, dithiothreitol; h-s-n, heptamer-spacer-nonamer; TG, transgenic reporter construct.


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