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Infection and Immunity, December 1999, p. 6643-6651, Vol. 67, No. 12
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Peptide Localization and Gene Structure of Cryptdin 4, a Differentially Expressed Mouse Paneth Cell alpha -Defensin

Andre J. Ouellette,1,2,* Dalila Darmoul,3 Dat Tran,1,2 Kenneth M. Huttner,4 Jun Yuan,1 and Michael E. Selsted1,2

Departments of Pathology1 and Microbiology and Molecular Genetics,2 College of Medicine, University of California, Irvine, California 92697; Neuroendocrinologie et Biologie Cellulaire Digestives, INSERM U410, Faculté de Médicine Xavier Bichat, 75018 Paris, France3; and Section on Neonatology, Department of Pediatrics, Massachusetts General Hospital, Boston, Massachusetts 021144

Received 11 June 1999/Returned for modification 26 July 1999/Accepted 17 September 1999

Paneth cells in crypts of the small intestine express antimicrobial peptides, including alpha -defensins, termed cryptdins in mice. Of the known Paneth cell alpha -defensins, the cryptdin 4 gene is unique, because it is inactive in the duodenum and expressed at maximal levels in the distal small bowel (D. Darmoul and A. J. Ouellette, Am. J. Physiol. 271:G68-G74, 1996). With a cryptdin 4-specific antibody, immunohistochemical staining of ileal Paneth cells was strong and specific for cytoplasmic granules, demonstrating that this microbicidal peptide is a secretory product of Paneth cells in the distal small intestine. Consistent with the pattern of cryptdin 4 mRNA distribution along the length of the gut, the cryptdin 4 peptide was not detected in duodenum. Structurally, the cryptdin 4 gene resembles other Paneth cell alpha -defensin genes. Its two exons, transcriptional start site, intron, splice sites, and 3' flanking sequences are characteristic of the highly conserved mouse alpha -defensin genes. However, in the region upstream of the transcriptional initiation site, the cryptdin 4 gene contains a repeated 130-bp element that is unique to this alpha -defensin gene. Every independent cryptdin 4 genomic clone examined carries the repeated element, which contains putative recognition sequences for TF-IID-EIIA, cMyc-RS-1, and IgHC.2/CuE1.1; the repeat proximal to the start of transcription replaces DNA at the corresponding position in other mouse alpha -defensin genes. We speculate that this unique duplicated element may have a cis-acting regulatory role in the positional specificity of cryptdin 4 gene expression.

* Corresponding author. Mailing address: Department of Pathology, University of California College of Medicine, Irvine, CA 92697-4800. Phone: (949) 824-4647. Fax: (949) 824-1098. E-mail: aouellet{at}uci.edu.

Infection and Immunity, December 1999, p. 6643-6651, Vol. 67, No. 12
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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