SHORT COM M UNICATION The Human dead ringer/bright Homolog, DRIL1: cDNA Cloning, Gene Structure, and Mapping to D19S886, a Marker on 19p13.3 That Is Strictly Linked to the Peutz–Jeghers Syndrome R. Daniel Kortschak,* Heike Reimann,² Michael Zimmer,‡ Helen J. Eyre,§ Robert Saint,* ,1 and Dieter E. Jenne² * Department of Genetics, University of Adelaide, Adelaide, South Australia 5005, Australia; ² Department of Neuroimmunology, Max- Planck-Institute of Psychiatry, am Klopferspitz 18A, D-82152 Planneg Martinsried, Germany; ‡Institute of Clinical Biochemistry and Pathobiochemistry, University of Wu ¨rzburg, 97080 Wu ¨rzburg, Germany; and §Department of Cytogenetics and Molecular Genetics, Women’s and Children’s Hospital, North Adelaide, South Australia 5005, Australia Received October 20, 1997; accepted February 11, 1998 The Drosophila gene dead ringer (dri) was isolated as a novel gene encoding a sequence-specific DNA- binding protein. DRI is a founding member of a growing protein family whose members share a con- served DNA binding domain termed the A/T-rich in- teraction domain. dri is developmentally regulated, being expressed in a restricted set of cells including some neural cells and differentiating cells of the gut and salivary gland ducts. The mouse homolog of dri, bright, has been shown to be expressed in mature B-cells in the immune system, its product trans-ac- tivating expression through an IgH enhancer in transient transfection assays. We have cloned a hu- man dri/bright homolog, termed DRIL1. Here we re- port the exon–intron structure of the gene and show physical linkage within 80 kb to the D19S886 marker on 19p13.3. As this marker is intimately linked to the Peutz–Jeghers syndrome in several large pedigrees, human dri (DRIL1) is a candidate gene for this dis- order. © 1998 Academic Press Although a vast number of DNA-binding proteins have been described, they can be grouped into a small number of families on the basis of conserved DNA binding domains. Recently a new family of DNA binding proteins was identified (7, 9). This fam- ily contains a number of partially characterized genes of known or predicted regulatory significance, including the Drosophila dead ringer (dri) (7) and eyelid (15) genes; yeast SWI1 (11); the mammalian bright (9), jumonji (13), Smcx (2), and Smcy (1) genes; and the genes encoding the retinoblastoma binding factors, RBP1 and RBP2 (5). Two closely related members of this family, dead ringer and bright, have an extended region of similarity and encode proteins that bind to DNA, in vitro, with specificities similar to that of the Engrailed home- odomain protein (7, 9). The common domain in this group of proteins, the A/T-rich interaction domain (ARID), has been shown to be responsible for the DNA binding activity of these two members of the family (7, 9). Both dri and bright are implicated in developmental processes. Embryos lacking DRI show developmental defects, including segmentation and gut defects, and die during embryogenesis (Shandala et al., in preparation), while bright has been shown to be expressed in mature B-cells in the immune system, its product trans-activat- ing an IgH enhancer in transient transfection assays (9). There are several precedents for roles for homologs of Drosophila developmental regulatory genes in human genetic disease, e.g., PAX-3, implicated in Waardenburg syndrome (14), and patched, implicated in tumor forma- tion (8, 10). We sought to determine whether a human homolog of the Drosophila dri and mouse bright genes exists and, if so, whether it is a candidate for any mapped genetic developmental disorder. We report, here, the iso- lation, initial characterization, and localization of DRIL1, a human homolog of the Drosophila dead ringer and mouse bright ARID box genes. The initial sequence corresponding to DRIL1 was isolated using a degenerate PCR technique. Primers designed to be specific for extended ARID box se- quences were used to amplify fragments from a HeLa- derived Lambda ZAP cDNA library (Clontech). Two rounds of amplification with an annealing temperature of 37°C were performed: initially 30 cycles using the primers 5'-AARMGNAARGARTTYYT-3' and 5'-YKYT- CRCAYTCRTANGG-3', followed by 30 cycles using the primers 5'-CGGGATCCGAYYTNTTYWSNTTYATG-3' and 5'-GGAATTCGRTANARRTAYTTCATRTA-3'. The Sequence data from this article have been deposited with the GenBank Data Library under Accession No. U88047. 1 To whom correspondence should be addressed at the Department of Genetics, University of Adelaide, Adelaide, South Australia 5005, Australia. Telephone: 61-8-8303 5563. Fax: 61-8-8303 4399. E-mail: rsaint@genetics.adelaide.edu.au. GENOMICS 51, 288 –292 (1998) ARTICLE NO. GE985259 288 0888-7543/98 $25.00 Copyright © 1998 by Academic Press All rights of reproduction in any form reserved.